Method for making non-dehiscent sesame

A breeding method for a non-dehiscent sesame plant has been developed. Non-dehiscent sesame varieties are characterized by having sufficient capsule split, capsule opening, capsule placenta attachment, capsule constriction, and capsule membrane attachment to allow seed retention in the field after physiological maturity during adverse weather conditions such as rain, wind, and dew and also to allow ready release of seed from the capsule during mechanized harvesting with minimal broken seed. A mechanical method is also provided for quantitative screening of sesame plants for non-dehiscence.

TECHNICAL FIELD OF THE INVENTION 
This invention concerns sesame plant breeding and providing sesame plant 
varieties appropriate for mechanized harvesting. 
BACKGROUND OF THE INVENTION 
Sesame, or Sesamum indicum, is a tropical annual cultivated for its oil and 
its nut flavored seeds. The sesame plant grows to a height of 2-7 feet, 
and at its leaf axils are found capsules which contain the sesame seed. 
Upon maturity in nature, the capsules holding the sesame seeds begin to 
dry down, the capsules normally split open, and the seeds fall out. 
Commercially, the harvester tries to recover as much seed as possible from 
mature capsules. From ancient times through the present, the opening of 
the capsule has been the major factor in attempting to successfully 
collect the seed. Harvesting methods, weather, and plant characteristics 
all contribute to the amount of seed recovered. 
The majority of the world's sesame is harvested manually. With manual 
nonmechanized methods, it is desirable for the sesame seed to fall readily 
from the plant. Upon physiological maturity, the sesame stalks are cut, 
tied into small bundles, and then stacked in shocks. Further harvesting 
procedures vary from country to country and from area to area within 
countries. Some move the shocks to a threshing floor so that the seed that 
falls out can be recovered. Others put plastic or cloth in the fields 
under the shocks to catch the seed. For manual harvesting methods in which 
the dried, shocked sesame is moved to a threshing floor or over a plastic 
or cloth, preferred plant varieties include dehiscent, or super 
shattering, in which less than 10% of the seeds set are retained in the 
capsule. 
Other methods involve leaving the shocks in the fields, and when the shocks 
are dry, the sesame is turned upside down and struck with an implement to 
shake out all of the seed. For this type of manual harvesting method, it 
is preferred that the capsule hold as much of the sesame seed as possible 
until the farmer inverts the stalk. Plant varieties rated as shattering 
which retain as much seed as possible before inversion are preferred. 
Common methods of manual harvest are discussed in Weiss, E. A., (1971). 
Castor, Sesame, and Safflower. Leonard Hill Books, London, England. 
In an effort to mechanize the harvest of sesame, D. G. Langham introduced 
the use of swathers in Venezuela in 1944. The swathers were used to cut 
the sesame plants, manual labor was used to bundle and shock the cut 
plants, and combines were brought in to thrash the shocks. It was 
determined that seed shattering during mechanized harvesting methods 
caused considerable loss of sesame seed. While mechanization was 
considered to be essential for crop production in the Western hemisphere, 
it became obvious that the dehiscence of the sesame seed pod was the 
principal obstacle to the widespread acceptance of sesame as a commercial 
crop. (Langham, D. G. 1949. "Improvement of Sesame in Venezuela," 
Proceedings First International Sesame Conference, Clemson Agricultural 
College, Clemson, S.C., pp. 74-79). As programs to introduce sesame 
production in the United States in Arizona, South Carolina, Nebraska, 
Oklahoma, and Texas were initiated, mechanization was considered essential 
due to high labor costs. Kalton, one of the Texas researchers, reported 
that the shattering nature of available strains was the main obstacle in 
complete mechanization of the sesame crop. (Kalton, R. 1949. "Sesame, a 
promising new oilseed crop for Texas," Proc First International Sesame 
Conference, Clemson Agricultural College, Clemson, S.C., pp. 62-66). 
In 1943, D. G. Langham found a mutation on a sesame plant. Capsules did not 
open on plants expressing this mutation (FIG. 8). In succeeding 
generations, Langham showed that it was a recessive single gene which 
produced this indehiscence, where all the seeds were retained inside the 
unopened capsule. While it was believed that indehiscence would solve the 
problem of seed loss on mechanized harvesting, it was found that the 
capsules were too tough to effectively release the seed. Many of the 
capsules passed through a combine without opening. When more rigorous 
combining was attempted, an increase in efficiency of capsule opening was 
achieved but at the expense of seed quality. Seeds were broken due to the 
more rigorous combine conditions, and the broken seeds released free fatty 
acids. Chemical reactions with free fatty acids led to rancidity and 
concomitant undesirability of the harvested seed. 
The indehiscent sesame varieties described above were used by various plant 
breeders in an attempt to develop desirable sesame lines. In addition to 
traditional cross-breeding approaches, some attempted to alter the 
chromosome numbers through tetraploids and interspecific crosses. Yermanos 
attempted to improve release of seed by increasing the length of the 
capsule so that there would be more surface for the combine to crack the 
capsules open (personal communication). Unfortunately, even with a small 
opening on the top of the capsule, a high percentage of broken seed was 
found on mechanized harvesting, preventing commercial use of this sesame 
line. 
D. G. Langham reported in the late 1950's that the placenta attachment 
between each sesame seed and the placenta was important in the retention 
of seed in the capsule. He believed that he could improve the shatter 
resistance of sesame with increased placenta attachment but did not 
believe that all the seed could be retained in the capsule. However, 
Yermanos reported that during capsule maturity, the placenta attachment 
gradually weakens and is obliterated when the capsule is completely 
desiccated. (Yermanos, D. M. 1980. "Sesame. Hybridization of crop plants," 
Am Soc Agronomy-Crop Sci of America, pp. 549-563). Thus, it appeared that 
the placenta attachment would have little effect on seed retention in dry, 
mature capsules during harvesting. A seamless gene which retained all the 
seed in the capsules was discovered by D. G. Langham and D. R. Langham in 
1986 (FIG. 9). This was crossed with shattering types, and some progeny 
had an opening at the tip of the capsule. The seamless capsules were 
similar to the indehiscent capsules in that it was too difficult to remove 
the seed from the capsule without damaging the seed. 
In 1982, the first non-shattering line (retaining 50-70% of the seeds set) 
requiring no manual labor was introduced. This line could be harvested by 
swathing the sesame, leaving it to dry in the field, and then picking it 
up by a combine. Although complete mechanization was achieved, extensive 
loss of seed due to adverse weather conditions continued to occur. 
Other varieties were developed between 1988 and 1997 which allowed for 
direct combining with 70-90% seed retention, but extensive loss of seed 
due to wind and rain continued to occur. Lines that generally yielded 80% 
of the seed under ideal conditions would yield only 45-65% under adverse 
conditions. Thus, while many of the crosses began to moderate the 
deleterious effects of mechanized harvesting, none were able to increase 
the yields to the level of manually harvesting shattering cultivars. 
New lines of sesame have now been discovered which are defined by a new 
category of dehiscence: non-dehiscence. These lines retain most of the 
seed within the capsule despite adverse weather conditions such as wind 
and rain. The sesame lines of the present invention retain a sufficient 
amount of sesame seed during mechanized harvesting to be competitive with 
manual harvesting. The invention permits mechanized harvesting to be used 
with minimization of seed breakage because extensive combining is not 
required to obtain practical yields. Thus, the invention permits reduction 
of manual labor and concomitant economical and more rapid harvesting of 
sesame seed as a commercial crop. 
SUMMARY OF THE INVENTION 
In one aspect, the present invention relates to non-dehiscent sesame plants 
characterized by having greater than or equal to about 65% of the total 
amount of sesame seed retained in unharvested capsules subjected to the 
shaker test, less than or equal to about 10% of the total amount of sesame 
seed retained in mechanically harvested capsules, and less than or equal 
to about 3% of the total amount of sesame seed which is released from 
capsules broken during mechanical harvesting. 
In another aspect, the present invention relates to non-dehiscent sesame 
plants characterized by having greater than or equal to about 65% of the 
total amount of sesame seed retained in unharvested capsules subjected to 
the shaker test, less than or equal to about 5% of the total amount of 
sesame seed retained in mechanically harvested capsules, and less than or 
equal to about 7% of the total amount of sesame seed which is released 
from capsules broken during mechanical harvesting. 
In another aspect, the present invention relates to non-dehiscent sesame 
plants characterized by having greater than or equal to about 65% of the 
total amount of sesame seed retained in unharvested capsules subjected to 
the shaker test, less than or equal to about 5% of the total amount of 
sesame seed retained in mechanically harvested capsules, and less than or 
equal to about 3% of the total amount of sesame seed which is released 
from capsules broken during mechanical harvesting. 
In another aspect, the present invention relates to seeds and progeny of 
non-dehiscent sesame plants. 
In yet another aspect, the present invention relates to a non-dehiscent 
sesame plant characterized by a capsule opening that is slightly to barely 
open, and a good to moderate capsule placenta attachment. 
In yet another aspect, the present invention relates to a non-dehiscent 
sesame plant characterized by capsule split, capsule opening, capsule 
membrane completeness, capsule constriction, capsule membrane attachment, 
and capsule placenta attachment. 
In yet another aspect, the present invention relates to a non-dehiscent 
sesame plant selected from the sesame lines Sesaco 22 (S22), Sesaco 23 
(S23), Sesaco 24 (S24), 19A, and 11W, representative seed of said S22, 
S23, S24, 19A, and 11W having been deposited under ATCC accession number 
PTA-1400, PTA-1401, PTA-1402, PTA-1399, and PTA-1398 respectively. 
In another aspect, the invention relates to seeds and progeny of 
non-dehiscent sesame plants derived from the sesame lines S22, S23, S24, 
19A, and 11W. 
In yet another aspect, the present invention relates to a non-dehiscent 
sesame plant having the same non-dehiscent phenotype as a plant selected 
from the sesame lines S22, S23, S24, 19A, and 11W. 
In yet another aspect, the present invention relates to seeds and progeny 
derived from a non-dehiscent sesame plant having the same non-dehiscent 
phenotype as a plant selected from the sesame lines S22, S23, S24, 19A, 
and 11W. 
In one aspect, the present invention relates to a method of breeding a 
non-dehiscent sesame by combining shatter resistant characteristics of 
capsule split, capsule opening, capsule membrane completeness, capsule 
constriction, capsule membrane attachment, and capsule placenta 
attachment. 
In another aspect, the present invention relates to seeds produced 
according to the sesame breeding method for non-dehiscence. 
In yet another aspect, the present invention relates to progeny plants 
produced according to the sesame breeding method for non-dehiscence. 
In yet another aspect, the present invention relates to a method for 
screening sesame plant varieties for non-dehiscence. 
In yet another aspect, the present invention relates to a sesame plant 
identified by the non-dehiscent screening test.

DETAILED DESCRIPTION OF THE INVENTION 
Novel non-dehiscent lines of sesame and methods for producing non-dehiscent 
lines have now been developed. In the method for producing non-dehiscent 
lines, the breeder following the teachings of the present invention may 
select appropriate sesame plants to cross with other sesame plants to 
result in progeny sesame plants having the desired characteristics of 
capsules to allow for better retention and less breakage of sesame seed 
during mechanized harvesting despite exposure of the sesame crop to 
adverse weather conditions such as rain and wind. These non-dehiscent 
lines provide for the first time a means by which commercially acceptable 
sesame seed can be mechanically harvested at a production rate comparable 
to manual harvesting regardless of weather conditions. 
Non-dehiscent sesame lines are a type of shatter resistant sesame 
identified by the amount of sesame seed retained in the mature, dry 
capsules. These lines must hold the sesame seeds in the mature, dry 
capsules on the plants in the field but must release the seeds within the 
combine as easily as possible without breakage of the seeds. As used 
herein, "broken" sesame seed is defined as large and small pieces of 
kernels of sesame seed which have been broken and which remain in the 
harvested sample after the removal of dockage (Standards for Inspection 
and Grading of Sesame Seed, Hudson Laboratories, Nov. 1, 1993). The amount 
of seed retention is influenced by diverse characters of mature capsules 
which are manipulated through the present inventive method of breeding and 
selection processes to achieve non-dehiscence. It has been found that 
sesame plants phenotypically expressing only one of these characters does 
not have enough shatter resistance to qualify as non-dehiscent. However, 
in the method of the invention, multiple phenotypic characters become 
expressed in progeny plants and these multiple phenotypic characters 
provide the progeny plants with the desired non-dehiscence. These 
characters with corresponding rating methodology are summarized in Table 
I. 
Capsule Characters Affecting Non-dehiscence 
Sesame seed is produced in capsules (FIG. 1) that are in the leaf axils of 
the plants. Each capsule consists of carpels, and each carpel normally has 
two locules or seed chambers. Most sesame in the Western Hemisphere is 
bicarpellate, but tricarpellate and quadricarpellate cultivars are found 
in Asia and Africa. There has been no attempt to add 
TABLE I 
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Capsule Shatter Resistant Characteristics Used in 
Development of Non-dehiscent Lines 
Characteristics 
Abbreviation of the capsule Rating/value 
System 
______________________________________ 
Capsule split 
extent of split between the 
no split to complete split; 
(TS) carpels exposing the scale of 0-8; 
membranes but not 1 = split almost to 
exposing seed base of the capsules, 
4 = split halfway 
down capsule; 
7 = barely split; 8 = no 
split; in capsules 
where there is a 
difference in split on 
each side of the 
capsule, the greater 
split measurement is taken 
Capsule extent of opening between no opening to 
opening (TO) the carpels with complete opening; scale 
membranes opening of 0-8; 1 = open 
enough to expose almost to bottom of 
seed and/or seed the capsule; 4 = open 
chamber halfway down; 7 = 
barely open; 
8 = no opening 
Capsule the amount of complete membrane 
membrane missing membranes to no membrane; 
completeness the carpels scale of 0-8; 
(TM) 0 = no membrane, 1 = most 
membranes incomplete, 
4 = half of 
membranes incomplete, 
7 = complete 
membranes, 
8 = membranes with no 
holes 
Capsule degree of constriction no constriction to 
good constriction; 
constriction of the capsule around scale of 0-8; 
1 = little seed, 4 = half the 
(TC) the seeds as shown seed, 7 = most of the seed 
by the amount of seed 
remaining in the capsule 
after the placenta 
is removed 
Capsule amount of separation large to small 
membrane between the membrane separation between the 
attachment and placenta membrane and placenta; 
(TA) scale 0-8; 
0 = no membrane, 
1 = large separation, 
4 = medium separation, 
and 7 = little 
separation 
Capsule strength of placenta no to good placenta 
placenta attachment attachment 
attachment scale of 0-8; 
(TP) 1 = minimal placenta 
attachment, 
4 = some placenta 
attachment, 
7 = good placenta 
attachment 
______________________________________ 
non-dehiscence to lines other than bicarpellate, but it is expected that 
non-dehiscent lines can be made in tricarpellate and quadricarpellate 
cultivars using this invention. Normally, there is one row of 15-22 seeds 
per locule. In the center of the capsule, there is a placenta that 
nourishes the seeds during growth through a placenta attachment. In a 
cross-section, the outer layer is the epidermis followed by multiple 
layers of mesocarp (rounded parenchyma cells) with the endocarp (heavily 
lignified sclerenchyma cells) surrounding the seeds. (Day, J. 1998. "The 
mechanism of indehiscence in sesame-features that might be useful in a 
breeding programme"; presented at Induced Mutations for Sesame 
Improvements meeting held Apr. 6-10, 1998 in Bangkok, Thailand). In 
splitting the capsule between carpels, the endocarp is between the seeds 
in the two carpels. This part of the endocarp is known as the membrane. 
Although there is no visible line on the epidermis between the carpels, 
the mesocarp layers are arrayed at a suture to allow a splitting between 
the carpels. The force of the splitting is provided by the drying of the 
mesocarp layers of the cells. 
The rating system used for providing values to the characters related to 
shatter resistance is a 0-8 scale. A subjective scale is used instead of 
measurements since the capsules vary considerably in length and width. The 
expression of the character will also vary between capsules on the same 
plant and capsules on different plants. The 0-8 scale provides a 
subjective method of averaging observations in order to differentiate 
varying levels of shatter resistance. The ratings can be used to compare 
lines within a nursery in a given year, but do not necessarily apply 
across different locations and/or years. However, the relationships 
between lines remains constant across different locations and years. FIGS. 
2A-2C, 3A-3C, 4A-4C, 5A-5C, 6A-6C, and 7A-7C show three levels of ratings 
for each character. 
Capsule Split ("TS") 
One of the characters of sesame found to be useful in breeding 
non-dehiscent sesame is related to splitting of the capsule which contains 
the sesame seeds in the mature plant (FIG. 2A-2C). Regardless of the 
degree of shatter resistance, most sesame lines follow the same sequence 
of capsule maturity. As a capsule matures, it begins to dry down, turning 
brown in the process. Usually after the capsule is brown, the tip of the 
capsule splits along the suture that eventually separates the carpels. The 
amount of capsule split is greatly affected by weather. In nature, 
capsules exhibiting characteristic capsule split will be open under dry 
conditions. During a heavy dew or rain, the capsules will close. If a fog 
or drizzle persists during the day, the capsules will remain relatively 
closed, but in the sun, they will open to a similar extent as found under 
dry conditions. If the dew point is high, the splitting of the capsules is 
accelerated as each opening appears to increase the rupturing along the 
suture between the carpels. When the dew point is low, there is less 
splitting of the capsule. A capsule may show a different rate of capsule 
split on each side of the capsule, and in this case, the greater split 
measurement is taken. 
A desirable phenotypic character utilized in the breeding of non-dehiscent 
sesame plants is capsule split for both sides from the top of the capsule 
to approximately the base of the capsule, or TS1 (FIG. 2a). This rupturing 
of the epidermis and mesocarp is critical in making it easier to thresh 
the capsule in the combine. In lines where the split is not complete, the 
threshing must be more aggressive to remove the seed from the capsule. 
This additional aggressiveness can break the seed. 
Capsule Opening ("TO") 
Another character found to be important in selection of candidate plants 
for breeding is capsule opening. As the capsule splits the epidermis and 
mesocarp layers of cells along the suture, the carpels can open between 
the membranes, exposing the seed. The level of opening between these 
membranes is shown in FIG. 3A-3C. It is desirable to select candidate 
breeding plants with a TO character between TO6 and TO7 according to the 
scale set forth in Table I. TO6 and TO7 are defined as slightly open to 
barely open, respectively. It is desirable for the non-dehiscent lines to 
exhibit an opening at the top of the membrane, ranging from moderate to 
barely open, which generally reveals only the top two to three seeds in 
each locule. The capsule opening acts as a vent for moisture and to allow 
a starting point for the combine to open the capsule for release of the 
seed. At plant physiological maturity, the capsules in the center of the 
capsule zone have approximately 51-60% moisture which must be reduced 
substantially before harvest. Without an opening, moisture has to escape 
by migrating through the capsule wall. With an opening, the moisture can 
escape through the tip. The advantage of capsules with an opening can be 
seen in that a field of indehiscent or seamless sesame ready for harvest 
cannot be harvested until five to seven days after a light rain, whereas a 
field of sesame with an opening in the capsule can be harvested within one 
to two days following a light rain, allowing for less exposure to adverse 
weather conditions. Further, while indehiscent or seamless lines require 
that the capsule be broken across the locule for the seed to be released, 
having a capsule opening allows the seed to escape from the capsule 
without first breaking the capsule, thus assisting in mechanized 
harvesting. There are indehiscent and seamless lines with a slight opening 
at the top. However, the indehiscent and seamless lines do not have the TS 
to the base of the capsule and thus do not open as readily in the combine. 
Capsule Membrane Completeness ("TM") 
Another character to be evaluated in candidate plants to be used in the 
method of the invention is capsule membrane completeness. In some known 
lines of sesame, there are no membranes between the carpels, and the 
absence of membranes allows seed to fall out when the capsule opens. In 
some lines, portions of the membrane are missing as shown in FIGS. 4A-4B. 
Capsule membrane completeness is preferably TM7, or complete membrane, in 
non-dehiscent lines. All known lines of sesame have an opening in the top 
of the membrane which allows the seed to slip out of the capsule. This 
hole must be present or the threshing will have to be too aggressive. With 
the exception of membraneless lines, the parts of the membrane missing 
varies from capsule to capsule within the plant. This rating is based on 
opening ten capsules and averaging the missing membranes. 
Capsule Constriction ("TC") 
Another character to be evaluated in candidate plants to be used in the 
method of the invention is capsule constriction. The degree of 
constriction of the capsule around the seeds prevents seed from falling 
out of the capsule through the capsule opening. This characteristic is 
measured as capsule constriction. When the carpels are separated, the 
placenta (the part of the capsule the seeds are attached to through 
placenta attachment as shown in FIG. 7D) will go with one carpel or the 
other. Constriction can be determined in the carpel without the placenta 
and thus independent of placenta attachment. This carpel can be inverted 
and the amount of seed remaining in the capsule determines the amount of 
constriction as shown in FIGS. 5A-5C. The desirable range for capsule 
constriction is TC3 to TC5, or moderate seed retention. Ratings of TC6 to 
TC8 make it difficult to get the seed out of the capsule in the combine. 
Ratings of TC0 to TC2 are associated with capsules that allow the seed to 
rattle in the base and apply pressure to the upper seed, thus breaking the 
placenta attachment. 
Capsule Membrane Attachment ("TA") 
Another character to be evaluated in candidate plants to be used in the 
method of the invention is capsule membrane attachment. This is a 
characteristic having a high correlation with non-dehiscent lines. When 
the capsule is opened, most non-dehiscent lines have a membrane that 
extends adjacent to the placenta. By adjacent, it is meant that a rating 
of moderate to little separation between the membrane and placenta is 
observable. More specifically, when the carpels in a capsule are 
separated, the edges of the membrane can pull away from the placenta. The 
rating is, therefore, taken by restoring the membrane to its natural 
position as shown in FIGS. 6A-6C, so that any folds, curls, or distortions 
of the membrane caused by opening the capsule are eliminated prior to 
taking the rating. Non-dehiscent lines have preferred ratings of TA5 to 
TA8, or moderate to little separation between the membrane and placenta. 
Placenta Attachment ("TP") 
Another character to be evaluated in candidate plants to be used in the 
method of the invention is attachment of the seed to the placenta through 
a placenta attachment. During growth, all sesame seeds are attached to the 
placenta through a placenta attachment. At physiological maturity of the 
seed, the placenta attachment will dry. There are lines where the dry 
placenta attachment will keep the seed attached to the placenta, and other 
lines, where the placenta attachment breaks under normal drydown 
conditions as shown in FIGS. 7A-7C. The level of placenta attachment is 
the most critical aspect of non-dehiscent sesame. The TP rating needs to 
be between TP6 and TP8, or moderate to good capsule placenta attachment at 
the top of the capsule, but the higher the rating, the better. When most 
capsules first open, the rating is often TP7 to TP8, but within a few days 
with even slight breezes and rubbing of capsules against neighboring 
plants or other capsules, on most lines, the placenta attachment breaks 
down and the seeds begin to fall out of the capsules. Unlike the placenta 
attachment described by Yermanos as weakened and obliterated in dry 
capsules, the preferred placenta attachment does not weaken on dry down. 
The ideal placenta attachment is one that will hold the seed in the 
capsules even for an extended period during bad weather in terms of wind 
and moisture (dews or rain). However, the placenta attachment must be weak 
enough to release the seed once the capsule is inside the combine. To 
date, no lines with too much placenta attachment to prevent release of the 
seed within the combine are known. 
In some lines, there is a difference in the amount of placenta attachment 
within the capsule. Good placenta attachment at the base of the capsule is 
not essential when there is good attachment at the top of the capsule. The 
seed held at the top blocks the bottom seed from exiting the capsule. 
Interrelationships between Capsule Characters 
On all six characters, better ratings of one can offset lower ratings on 
other characters. There is a small allowance between lines that hold 
enough and lines that hold too much. The plants must hold the seeds until 
in the combine but must release the seeds within the combine as easily as 
possible with as little breakage of seeds as possible. 
TS and TO 
It has been found that with high TS ratings, the capsules do not open 
readily in the combine, requiring more aggressive threshing. The adhesion 
between the membranes is essential. Non-dehiscent sesame needs high TO 
ratings and low TS ratings. 
TO and TM 
Generally, there is a correlation between TM and TO in that if there are 
parts of the membranes missing, there is less adhesion between the 
carpels, and the TO rating will be low. Thus, although high TM ratings do 
not necessarily lead to high TO ratings, low TM ratings are usually 
associated with low TO ratings. 
TO and TP 
A high TP rating can offset a lower TO rating. However, a high TO rating 
will not offset a lower TP rating. 
TO and TA 
The added membrane surface on a high TA rating provides more adhesion 
between the membranes. This allows the capsule to split at the epidermis 
and mesocarp but not open to expose the seed. Thus, although high TA 
ratings do not necessarily lead to high TO ratings, low TA ratings are 
usually associated with low TO ratings. 
TC and TA 
A higher TA rating is usually associated with a high TC rating. The space 
between the membrane and placenta should be just large enough to provide 
sufficient volume around the seeds to reduce constriction. When TA=8, 
there is too much constriction. 
TC and TP 
Higher TC ratings can offset lower TP ratings. There are lines that hold 
enough seed to qualify as non-dehiscent through TC alone, but these lines 
do not release the seed in the combine. Logically, if the TP is high 
enough, and in combination with TO, the top of the capsule is blocked from 
releasing seed, no constriction would be necessary. However, to date, all 
non-dehiscent lines have some constriction. 
Methodology for Developing Non-Dehiscent Sesame 
By incorporating the above-identified shatter resistant characters into 
commercially suitable sesame lines, non-dehiscent sesame lines have been 
developed. The starting point for developing non-dehiscent sesame is 
acquisition of lines which have the shatter resistant characters. 
Representative sources for sesame lines necessary for the development of 
non-dehiscent sesame lines include the National Seed Storage Laboratory 
(NSSL) in Ft. Collins, Colo., and the Plant Genetic Resources Conservation 
Unit (S9) in Griffin, Ga. These collections were also deposited with the 
Food and Agriculture Organization of the United Nations (FAO) sesame 
collections maintained in South Korea and Kenya. In order to ensure the 
purity of these lines, one can grow out the materials for a year before 
selecting initial parent plants with required capsule characters. Table II 
provides representative sources for the different capsule characters 
required for developing non-dehiscent sesame, and reference to specific 
lines and crosses in the following discussion are made by SID codes. 
The line 111 has the best TO, but it is offset by not having enough TS. The 
TO in 118, 700, 701, and 702 is not good enough for non-dehiscence, but 
crosses between these four lines and 111 are the fastest avenue to the 
preferred amount of TS and TO. The line 111 has too much TC, while 118, 
700, 701, and 702 do not have enough TC. Crosses between these four lines 
and 111 are the fastest avenue to the preferred amount of TC. Crosses 
between 118, 700, 701, and 702 and 111 also provide the preferred level of 
TA and TM. 
TABLE II 
______________________________________ 
Sources for Capsule Characters Used in 
Development of Non-dehiscent Sesame 
Character SID.sup.a 
SESAN.sup.b 
Sesame PI Number.sup.c 
______________________________________ 
Capsule split (TS) 
118 T118 PI 426944 
700 T700 PI 292144 
701 T701 PI 292145 
702 T702 PI 292146 
Capsule opening 118 T118 PI 426944 
(TO) 111 T111 PI 173955 
700 T700 PI 292144 
701 T701 PI 292145 
702 T702 PI 292146 
Capsule membrane 118 T118 PI 426944 
completeness (TM) 111 T111 PI 173955 
700 T700 PI 292144 
701 T701 PI 292145 
702 T702 PI 292146 
Capsule constriction (TC) 111 T111 PI 173955 
Capsule membrane 700 T700 PI 292144 
attachment (TA) 701 T701 PI 292145 
702 T702 PI 292146 
Placenta attachment (TP) ACE TACE PI 320959 
191 T191 .sup.d 
192 T192 .sup.d 
193 T193 .sup.d 
______________________________________ 
.sup.a Significant lines were given three character sesame identifiers 
(SID). 
.sup.b When lines were aquired from the NSSL and S9 collections, PI 
numbers were converted to Sesanumbers (SESAN). 
.sup.c NSSL and S9 identifying codes. 
The capsule architecture in terms of length and width of the 
above-identified five lines is similar in size enough to allow for quick 
selection of a line with the proper amount of TS, TO, TM, TC, and TA. In 
making these selections, leaving the plants in the weather for at least a 
month after drydown is essential, since the TO can break down over time 
and thus prevent accurate measurement of weather resistant TO. When 
looking at the 118, 700, 701, and 702 capsules at drydown, it might appear 
that the crosses with 111 are not necessary, but they are essential, since 
111 either provides the correct amount of adhesion or a reduction in the 
layers of the mesocarp that allows less opening force. 
Having strong enough TP is essential in developing non-dehiscence. The 
lines 111, 118, 700, 701, and 702 lines do not have sufficient TP to 
provide non-dehiscence. Therefore, preferably crosses are made between 
these lines and lines which have strong TP, e.g., both the 191/192/193 
(representing herein "191, 192, or 193") U.S. lines and the ACE Venezuelan 
line (equivalent to G8 in FIG. 10, FIG. 11, and FIG. 12). Less preferred 
is a method crossing with just one of the groups 191/192/193 or ACE. 
Preferably, the 191/192/193 group is utilized. 
In terms of the importance of the characters, the placenta attachment (TP) 
and capsule opening (TO) are the two most important characters for seed 
retention, and the capsule split (TS) and capsule constriction (TC) are 
the most important characters for seed release in the combine. 
Crossing of sesame may be done using standard techniques as generally 
outlined below. Sesame is generally self-pollinated. Flowers develop and 
mature from the base of the plant to the top. Each morning, flowers open, 
and shortly before or after opening, the anthers burst longitudinally, 
releasing the pollen. Simultaneously, the lobes of the stigma open and 
receive large quantities of pollen. In order to cross sesame, the flowers 
must be emasculated prior to pollination. Most sesame flowers have a fused 
corolla with the stamens attached to the corolla. The evening before the 
flowers are to open, the corolla (with stamens) is removed from the female 
flower. Similarly, the corolla is removed from the male flower and stored 
in a container overnight. The next morning, the pollen from the male 
flower is put onto the stigma of the female flower. There are many 
variations in specific techniques as detailed in Langham, D. G. 1944. 
"Natural and controlled pollination in sesame," J Heredity 35:254-256; 
Yermanos, D. M. 1980. "Sesame. Hybridization of crop plants," Am Soc 
Agronomy-Crop Sci of America, pp. 549-563; and Osman, H. E. 1985. "Studies 
in sesame: Hybridization and related techniques," FAO Plant Production and 
Protection Paper No. 66, pg. 145-156. Each cross provides 40 to 80 seeds 
and usually a minimum of two flowers are available to cross each day. The 
F1 plants can be grown in a greenhouse since selection for capsule 
characters needs to be done in the F2 plants and not the F1 plants. The 
greenhouse does not provide an adequate environment to select for 
preferred capsule characters in any generation beyond F1, since wind is 
not a factor and there is not enough variability in moisture in terms of 
humidity, rain, fog, and dew in a greenhouse. 
Preferably, the shatter resistant characters are aggregated into one line 
and then this line is crossed against commercially suitable sesame lines. 
In crossing non-dehiscent sesame lines against commercially acceptable 
shattering lines, there is no shatter resistance in the F1 plants. In most 
crosses, there will be individual F2 plants with enough shatter resistance 
to be tested for non-dehiscence. The larger the F2 population, the greater 
the chance of finding non-dehiscent F2 plants. 
Under ideal conditions, in Year 1, crosses between 111 and 118/700/701/702 
(representing herein "118, 700, 701, or 702") are made in a 
greenhouse/winter nursery to the F1 plants, grown in a greenhouse/winter 
nursery; and the F2 plants, grown in the field. The F2 plants are examined 
to find one or more plants having appropriate TS, TO, TM, TC, and TA 
characters, providing Stage 1 selections. In Year 2, Stage 1 selections 
are crossed with ACE/191/192/193 in a greenhouse/winter nursery; the F1 
plants, grown in a greenhouse/winter nursery; and the F2 plants, grown in 
the field. The F2 plants are examined to find one or more plants having 
appropriate TS, TO, TM, TC, TA, and TP characters, providing Stage 2 
selections. By the end of Year 2, at least one of the Stage 2 selections 
is non-dehiscent. Although it is possible to find F2 plants with 
non-dehiscence, most non-dehiscent F2 and F3 plants segregate lower levels 
of shatter resistance, and most pure non-dehiscent lines have been 
selected at the F4 level. Moreover, if the plants for making the initial 
crosses are obtained in the spring, an additional half year is necessary, 
since it is essential that the F2 plants are grown in the field so that 
non-dehiscence is measurable upon exposure to adverse weather conditions. 
In order to increase the odds of success, all permutations are crossed 
except the intercrossing within the following two groups: 118/700/701/702 
and 191/192/193. Each line is used as both male and female. In other 
words, the following crosses are made: 
EQU 111.times.118, 111.times.700, 111.times.701, 111.times.702, 111.times.ACE, 
111.times.191, 111.times.192, 111.times.193, 
EQU 118.times.ACE, 118.times.191, 118.times.192, 118.times.193, 
EQU 700.times.ACE, 700.times.191, 700.times.192, 700.times.193, 
EQU 701.times.ACE, 701.times.191, 701.times.192, 701.times.193, 
EQU 702.times.ACE, 702.times.191, 702.times.192, 702.times.193, 
EQU ACE.times.191, ACE.times.192, and ACE.times.193. 
The reciprocal crosses should also be made, i.e., 118.times.111, 
700.times.111, 702.times.111, etc. This is a total of 54 crosses. Doubling 
the crosses using different parent plants improves the odds of success. 
Selections of the F2 plants having the preferred combination of shatter 
resistant characters are made, and then a crossing plan is designed based 
on the combinations found in the F2 plants. Again, a large number of 
crosses (about 100-250 crosses) are made to increase the odds of success 
in producing the proper combination of genes. Preferably, crosses of ACE, 
191, 192, and 193 against the F1 plants of 111.times.118 700 701 702 ACE 
191 192 193 are made to provide the potential of an earlier F2 population 
that would have the appropriate mixture of shatter resistant characters. 
However, large populations (about 5,000-30,000 plants) of the latter 
scheme must be grown out. 
The genetic controls for the shatter resistant characters have not yet been 
determined. However, given the percentage of F2 plants that segregate with 
the proper combinations of characters, more than one gene is involved in 
each character. While there are no genetic maps of sesame, the fact that 
all of the shatter resistant characters have been moved together in many 
different paths indicates that none of the genes are on chromosomes in 
such a way that cross overs are necessary for success. Although the genes 
responsible for non-dehiscence have not yet been established, it is 
believed that once these genes are isolated, it will be possible to inject 
these genes into cells or protoplasts taken from sesame plants lacking 
non-dehiscence, thus producing from these injected cells or protoplasts 
plants which exhibit non-dehiscence. 
Crosses between non-dehiscent lines and other commercially suitable lines 
are providing selectable non-dehiscence in the F2 plants. In some cases, 
the F2 selections are pure for non-dehiscence. Thus, the preferred method 
of producing commercially acceptable, non-dehiscent sesame is to make 
non-dehiscence the first criteria for selection and then move 
non-dehiscence to commercially suitable lines. 
The present invention relates to non-dehiscent sesame seed and the sesame 
plant produced therefrom. It also relates to seeds and plants produced by 
crossing each non-dehiscent variety with itself, another non-dehiscent 
variety, or other sesame varieties. Sesame lines S22, S23, S24, 19A, and 
11W (representative seed having been deposited under ATCC accession number 
PTA-1400, PTA-1401, PTA-1402, PTA-1398, and PTA-1399, respectively) are 
exemplary non-dehiscent sesame lines which have been found to be readily 
reproducible over successive planting seasons. Unless otherwise stated, as 
used herein, the term plant includes plant cells, plant protoplasts, plant 
cell tissue cultures from which sesame plants can be regenerated, plant 
calli, plant clumps, plant cells that are intact in plants, or parts of 
plants, such as embryos, pollen, ovules, flowers, capsules, stems, leaves, 
seeds, roots, root tips, and the like. Further, unless otherwise stated, 
as used herein, the term progeny includes plants derived from plant cells, 
plant protoplasts, plant cell tissue cultures from which sesame plants can 
be regenerated, plant calli, plant clumps, plant cells that are intact in 
plants, or parts of plants, such as embryos, pollen, ovules, flowers, 
capsules, stems, leaves, seeds, roots, root tips, and the like. 
Tissue culture of sesame is currently being practiced in Korea, Japan, Sri 
Lanka and United States. It is possible for one of ordinary skill in the 
art to utilize sesame plants grown from tissue culture as parental lines 
in the production of non-dehiscent sesame. Further, it is possible to 
propagate non-dehiscent sesame through tissue culture methods. 
EXAMPLE 1 
Breeding Program for Non-Dehiscent Sesame 
Non-dehiscent sesame was developed by crossing lines with various shatter 
resistant characteristics. Over thirty-seven lines have been developed and 
tested. FIGS. 10-12 demonstrate three different avenues by which 
non-dehiscent lines were developed. In reading the diagrams from right to 
left, the initial SIDs can be seen. 
The sources in Table II are not non-dehiscent, and also not suitable to be 
used as commercial varieties in the United States. These sources were 
crossed against lines having commercially preferred characters such as 
light color and disease resistance. Commercially acceptable, non-dehiscent 
lines were obtained. It was determined that with the exception of G8 (a 
sister selection of ACE), none of the lines in FIG. 10-FIG. 12 not listed 
in Table II contributed to non-dehiscent capsules. 
Methodology for Measuring Non-dehiscence 
Non-dehiscent sesame are characterized by sufficient retention of seed 
within the capsules while in the field and ready release of the seed from 
the capsules during mechanized harvesting with minimal breakage of the 
seed. Methods for measuring the retention of seed in capsule in the field, 
release of seed from the capsule during harvest, and breakage of seed 
during harvest have been developed. 
Subjective Shatter Resistance Screening Measurement 
The crossing methodology described above results in thousands of 
segregating plants, and an initial screening method has been developed for 
selecting plants to be submitted for a more stringent, objective 
measurement of shatter resistance as an indicator of non-dehiscence. This 
screening method for measuring shatter resistance is a rating system based 
on the two parameters outlined in Table III below. Based on Table III, 
FIGS. 5A-5C illustrate the amount of seed present in the dry capsules for 
TI and KE. FIG. 5A represents TI1 and KE1; FIG. 5B represents TI4 and KE4; 
and FIG. 5C represents TI7 and KE7. 
TABLE III 
______________________________________ 
Parameters Used in Rating Shatter Resistance 
Name/ 
Abbreviation Measurement Rating/value System 
______________________________________ 
Upright seed 
amount of seed scale of 0-8; 
retention (TI) present in the dry 1 = at the bottom of the 
capsule with the capsule, 4 = halfway 
capsule still up the capsule; 
remaining on the 7 = at the 
plant, measured tip of the capsule; 
by how close the seed 8 = indehiscent or 
is to the tip seamless capsule 
of the capsule 
Inverted seed amount of seed present scale of 0-8; 
retention (KE) scale of 0-8; 1 = at the bottom of the 
capsule after the capsule, 4 = halfway 
capsule is gently up the capsule; 
removed from the 7 = at the tip 
plant, inverted, of the capsule; 
and twirled 8 = indehiscent or 
seamless capsule 
______________________________________ 
The TI and KE parameters have been combined into a two digit hold (HLD) 
rating. By this method of shatter resistance measurement, non-dehiscence 
is preferably defined by a hold rating with TI6-TI7 and KE6-KE7, i.e., 
HLD=66 or above. Hold ratings may deteriorate through various stages: 
first stage=when plant is first dry; second stage=fourteen days after 
drydown with no adverse weather conditions such as wind and/or rain; third 
stage=one week after exposure to adverse conditions; and fourth stage=one 
month after drydown and exposure to adverse weather conditions. 
Non-dehiscent sesame demonstrates a HLD rating of 66 or above at all four 
stages. This screening method is limited in that while it estimates the 
amount seed retention (TI) and seed release (KE), it does not provide an 
objective measurement of the amount of seed release and seed breakage 
during mechanized harvesting. 
Objective Shatter Resistance Measurement 
Since all of the capsule character ratings presented herein are subjective 
and can be dependent on the time of observation in relation to complete 
drydown, shatter resistance cannot be objectively measured directly from 
capsule character ratings. Since the screening method described above 
provides only a subjective measurement of shatter resistance, objective 
methodologies have been developed. 
Seed Retention 
Examples 2 and 3 give methods for determining the first requirement of 
non-dehiscence, i.e., retention of seeds in the capsule. 
EXAMPLE 2 
Seed Retention Measurement by Natural Means 
One method for measuring shatter resistance to determine non-dehiscence 
involves evaluating retention of seed in capsules before and after 
exposure to adverse weather conditions. Testing fields should be located 
where the sesame will be subject to wind and rain after drydown. 
Initially, representative lots of ten capsules were removed from different 
lines in a sesame nursery during a normal harvest at initial drydown while 
the capsules were still holding all of their seeds, i.e., HLD=77. The 
weight of the seeds in each lot established an average baseline weight of 
seeds per ten capsules. Three months after initial drydown and exposure to 
inclement weather, a second representative lot of ten capsules was taken 
from the same plot. The average weight of seed retained in ten capsules in 
the second round was divided by the average baseline weight from the first 
round to determine the percentage of retention, providing a measurement of 
weather shatter resistance. 
By this method of measuring shatter resistance, non-dehiscent lines have 
from about 65% to about 97% retention of seed after exposure to adverse 
weather conditions. While this method has the disadvantage of leaving the 
sesame in the field for extended periods after drydown, it can be used in 
remote areas where more technical methods are not possible. 
EXAMPLE 3 
Mechanized Seed Retention Measurement 
While the weather shatter resistance method in Example 2 is one method for 
measuring shatter resistance to determine non-dehiscence, it is generally 
not practical to leave a sesame nursery for over three months past harvest 
to measure seed retention. Consequently, a methodology using a machine has 
been developed to simulate the weathering of capsules. 
After examining the shaking action of several machines, a reciprocal shaker 
was chosen to simulate weather conditions in the determination of sesame 
seed retention rates. 
Some of the capsules at initial drydown were used to calibrate the shaker. 
A Lab-line Reciprocal Shaker (Lab-line Instruments, Inc., Melrose Park, 
Ill.) was set at a stroke length of 1.5 inches and 250 strokes per minute. 
Ten capsules were placed into a 250 ml Erlenmeyer flask and shaken for 
five minutes, and loose seed was removed and weighed. The capsules were 
shaken for an additional five minutes, and the loose seed was removed and 
weighed. The capsules were shaken for an additional five minutes, and the 
loose seed was removed and weighed. The seed retained in the capsules was 
removed and weighed. 
The weights from the shaker test were then compared to weights obtained by 
the weathering procedure given in Example 2. It was determined that there 
were differences between the shaker retention rate and the natural 
weathering retention rate in the 5, 10, and 15 minute tests, but that ten 
minutes was the best fit with the lowest differences between the shaker 
shatter resistance and the weather shatter resistance. Retention rates of 
samples shaken for 5 minutes were higher than the retention rate of the 
weathered samples. Retention rates of samples shaken for 15 minutes were 
lower than the retention rates of the weathered samples. Some samples 
shaken for 10 minutes had lower retention rates than those for weather 
samples, and other samples shaken for 10 minutes had higher retention 
rates than those for weather samples. Retention rates for samples shaken 
for ten straight minutes were compared to retention rates for samples 
shaken for two five-minute increments. There were no significant 
differences between the two groups. 
The preferred method for measuring seed retention in sesame as a 
measurement of non-dehiscence is summarized as follows. Ten capsules from 
the center of the capsule zone are harvested when the plants have 90-95% 
dry capsules, typically within about two weeks of initial drydown. Only 
capsules with all their seed are chosen. Only lines that have 95% of their 
capsules holding all the seed should be tested. If the plot is segregating 
hold, it should not be a candidate for non-dehiscent testing. On lines 
having a single capsule per leaf axil, two capsules are taken from five 
plants in the middle of the capsule zone. On lines having triple capsules 
per leaf axil, two capsules from the same leaf axil are taken from five 
plants. The plants should be in normal populations in the center of the 
rows or more than half a meter from end plants or plants at the edge of a 
gap, and should not be from a row on the edge of a planting block. The 
capsules are then dried down using heat lamps or ovens. The capsules are 
placed in a 250 ml Erlenmeyer flask and submitted to the shaker test, with 
a Lab-line reciprocal shaker preferably at a stroke length of 1.5 inches 
and run at 250 strokes per minute for 10 minutes. The seed released from 
the capsules is removed from the flask and weighed. The seed retained in 
the capsules is threshed, removed from the flask, and weighed. Shaker 
shatter resistance (SSR) equals the weight of the retained seed divided by 
the sum of the released and retained seed multiplied by 100 equals the 
percent seed retention. The procedure should be repeated a minimum of four 
more times. Statistically, the lowest and highest SSR should be discarded 
and the other values averaged to determine the line SSR. Non-dehiscent, 
indehiscent, and seamless sesame retain about 65% or more of the seed by 
this method. 
Other types of shakers may be used to perform the shaker test. However, the 
stroke length, shaking speed, and shaking time must be calibrated for each 
type of shaker to determine the parameters which correlate to the 
retention rate of weathered samples. 
While seed retention can be measured by the weathering method of Example 2 
or the shaker method of Example 3, the shaker method has significant 
advantages over the weathering method. The shaker method provides quicker 
results since the sesame does not have to be exposed to extended 
weathering periods. The weathering method may give inconsistent results 
from one crop to another since weather patterns will vary from one season 
to another and from one location to another. In contrast, the shaker 
method is not weather dependent and provides more consistent results. 
Seed Release/Breakage During Mechanized Harvesting: Thresh Yield Tests 
To test for the second requirement of non-dehiscent sesame, i.e., high 
yield during mechanized harvesting, Examples 4 and 5 provide methods by 
which seed release and seed release/breakage, respectively, are 
determined. 
EXAMPLE 4 
Plot Thresher Screening Method 
Plot threshers can be used to screen for capsule seed retention vs. seed 
yield rates. Capsules taken from the plot thresher are opened and examined 
for seed retention. The capsules from non-dehiscent lines have about 90% 
of the seed removed from the capsule by the thresher, while homozygous 
indehiscent lines, homozygous seamless lines, and lines with high TS 
and/or high TC retain more than 10% of the seed in the capsules. While 
plot threshers provide an indication of the seed yield rate, they do not 
provide a measurement of seed breakage during mechanized harvesting. The 
plot threshers are, thus, used to identify crops which are to be subjected 
to the more definitive combine test. 
EXAMPLE 5 
Combine Method for Measuring Seed Yield 
The preferred method for the thresh yield test measures the amount of seed 
released and the amount of seed broken during harvesting in a combine. A 
sesame crop of not less than ten acres having a seed moisture content of 
about 6% or less is selected for combining. The combine is set for the 
field conditions such that the seed is threshed as gently as possible. 
Generally, this means a low cylinder speed and wide open concaves. For 
example, a John Deere 9600 combine is adjusted to the lowest cylinder 
speed with the concave adjusted to the "corn" setting and air at the 
minimum setting; and while threshing, the concave is adjusted toward the 
"soy" setting until mature seeds are removed from the capsules. On an IHC 
1680 combine, initial settings are cylinder 350, air 450, and fine grain 
concave with wires in. Most sesame will have capsules at the top of the 
plant that do not contain mature seed, and this immature seed is not 
counted in these tests. Once the combine is set to obtain 99%-100% release 
of seed with broken seeds at less than or equal to 2%, the seed gathered 
during the setting process is dumped. 
To get a representative sample, the combining test is begun and continues 
until the bin is full up to the input auger. With a four foot probe, 
samples are taken from four locations in the combine bin, and the samples 
are co-mingled. While the combine is operating, at a minimum of 100 feet 
from the end of the field, five capsules are taken from each of twenty 
plants at positions ranging from the bottom to the top of the plant but 
not including the immature seed capsules at the very top. In the same 
area, a container such as an oil changing pan is thrown between the wheels 
of the combine as it passes to catch the capsules that have gone through 
the combine and are going over the top of the screens/sieves. This 
procedure is repeated four more times. In the laboratory, the seed is 
threshed out of the 100 capsules taken from the twenty plants, and the 
seed is weighed. At random, 100 representative capsules that came out of 
the combine are selected. Seed from these capsules are threshed out and 
weighed. The weight of seed retained in the capsules is divided by the 
weight of the 100 capsules taken prior to combining. The seed samples 
taken from the combine bin are thoroughly mixed and a 60 gram sample is 
taken. The seeds are separated into three groups: whole sesame seeds, 
broken seeds, and non-sesame chaff/foreign matter/immature seeds. After 
weighing the first two groups, the weight of the broken seeds is divided 
by the sum of the weights of the whole seeds and the broken seeds. 
In non-dehiscent lines, the capsules preferably retain less than or equal 
to about 10% of the seed during combining. Comparatively, indehiscent and 
seamless lines retain more than 10% of the seed during combining. More 
preferably, non-dehiscent lines have less than about 7% broken seeds by 
weight after combining, whereas indehiscent and seamless lines have more 
than 7% broken seeds. Thus, non-dehiscent lines are identified as having 
about 65% or more seed retention using the mechanical shaker test, about 
10% or less seed retention in the thresh yield test, and less than or 
equal to about 7% broken seed in the thresh yield test.