Development of Rice Mutant Variety with Higher Yield and Improved
Agronomic Traits through Carbon Ion Beam Irradiation
A. N. K. Mamun1, M. H. Kabir1, A. K. Azad2, M. A. Azam2, M. R. Islam1, M. T. Jahan1,
Md. Monirul Islam1, Pronabananda Das1 and Md. Monirul Islam1
1

Plant Biotechnology and Genetic Engineering Division, Institute of Food andRadiation Biology, Atomic Energy Research Establishment, Bangladesh Atomic
Energy Commission, Bangladesh,

2 Plant Breeding Division, Bangladesh Institute of Nuclear Agriculture, Bangladesh.

Summary
BINA dhan14 is becoming more popular to the farmers due to the advantage ofhigher yield, early maturity, long grain fine rice and less water requirement over parent
as it is cultivated in rain-fed condition. Advanced promising mutant line obtained from
BRRI dhan 29 was released as a variety called BINA dhan18 and BINA dhan19 wasdeveloped from NERICA10. These varieties were developed through carbon ion beamirradiation from the QST, Takasaki, Japan with kind cooperation of FNCA during theperiod of late 2013 to early 2018. The main criteria of these varieties are lodging
resistant with higher yield, long grain fine rice. BINA dhan14 is cultivated during Boroand Aus season, BINA dhan18 is moderate duration with higher yield Boro rice and
BINA dhan19 is highly drought tolerant and also cultivated round the year. BINA
dhan18 is about 100 cm tall and requires 148-153 days (seed to seed) to mature.
Maturity was observed 13-15 days earlier than that of the parent variety BRRI dhan29.
Thousand grain weight is about 26.5 g, amylose and protein contents are 23.2% and7.06%, respectively. The average yield of 7.25 t/ha and the highest yield of 10.5 t/ha wasrecorded in favorable environmental condition. The average yield of BINA dhan19 isabout 3.84 t/ha and the maximum yield is 5.0 t/ha. The plant of BINA dhan19 is about
80-90 cm tall and does not get lodged. During severe drought the plant became almost
stop growing but when rain is started then it starts growing vigorously and gave almost
equal yield like favorable condition. It matures within 90-105 days and thousand grain
weights is about 23 g. The grain contains 7.32% protein and 23.8% amylose. Efforts arecontinuing towards indigenous rice cultivars of Lombur, B-11 and Hori dhan to develop
new mutant variety with higher yield, improved agronomic traits and tolerant to abiotic
stress.

1

Introduction
The continuous increase of population in the country demands more food. In turn,it requires more land for rice cultivation as rice is the staple food for 166 million
people in Bangladesh and also the most important food crop in Asia and rest of theworld (AIS, 2008). We are facing severe threat of climate change challenges in crop
sector. In Bangladesh, rice is grown three seasons in a year i.e., Aman season (July to
December), Boro season (December to May) and Aus season (March to June). Riceprovides 75% of total calories and 51% of protein in our population. About 60% of thetotal agricultural labor force is engaged for rice production (AIS, 2008). It covers
77.96% of total cropped area in Bangladesh (AIS, 2007) and contributing 14.6% of ournational GDP (BBS, 2004). It is a diploid species, with a small genome in comparison toother cultivated cereals (Moin et al., 2017). The average rice yield in the country is
about 2.5 t/ha. It is very low yield comparing to other rice growing countries in theworld (Islam, 1989). The low yield performance of rice in the country is the effect of
the cultivation of low yield potential genotypes, drought, flash-flood and soil salinity.
Salinity is also one of the significant abiotic constraints to rice cultivation in the
coastal areas of Bangladesh. Salt stress brings about huge losses in worldwide
agricultural productivity (Moradi et al., 2003; Yu et al., 2016). In the world, about 400
million hectares of land are affected by high soil salinity and about 1 million hectaresof land are affected by high salinity in the coastal regions in Bangladesh (Karim et al.,1990). Salinity hindrances almost every aspect of the physiology and biochemistry ofplants and significantly reduces yield. A number of indigenous cultivars are prone tolodging but had some degree of tolerant to salinity and submergence. Its grains are
medium bold and contains 10.3% protein and 28.6% amylose (Kamruzzaman andAsaduzzaman, 2004). Mutation Breeding Project of FNCA aimed for breeding
technology with major crops using irradiation either by gamma-ray or by ion-beam.
Mutations are the major source of genetic variability and artificial mutations can be
induced by mutagens (Wei et al., 2013; Oladosu et al., 2016). Many reports are available
in induced mutation of rice for genetic improvements with higher yield and otheragronomic traits (Azam and Uddin, 1999; Hidema et al., 2003). Recently, ion beamirradiation has been appeared to utilize an effective means of inducing mutations.
The biological effects of ion beams have also been investigated and observed to beshown a high relative biological effectiveness (RBE) in lethality, mutation and also totransfers high energy to the target compared to low linear energy transfer (LET)
radiation such as Gamma-rays, X-rays and electrons (Blakely, 1992). It has been also
demonstrated that ion beams induce mutations at high frequency and induce novel2

mutants in Arabidopsis (Hase et al., 2000, Shikazono et al., 2003, Tanaka et al., 1997,
2002). The use of ion beams for inducing mutations in rice (Oryza satva L.) breeding
has also been attempted (Hayashi et al. 2007; Hidema et al. 2003; Rakwal et al. 2008).
It is generally and widely accepted that in mutation breeding induced traits become
fixed during M2 to M4 generations (Azad et al., 2010; Hamid et al., 2006;
Shamsuzzaman et al., 1998; Azam and Uddin, 1999). Indigenous genotypes have been
indicated as the tremendous sources of genes for new alleles (Evenson and Gollin,
1997; Guevarra et al., 2001; Hoisington et al., 1999; Jackson, 1999; Tanksley and
McCouch, 1997). Based on the information mentioned above, the study was
undertaken to develop new mutant variety with higher yield and yield contributing
traits among the local rice genotypes in Bangladesh using carbon ion beam irradiation.
In rice, mutants were developed from immature embryos (Ookawa et al., 2014), calli
obtained from seeds (Serrat et al., 2014) and cell suspension cultures (Chen et al., 2013).
Seeds are, however, easier to handle and do not require a specialized structure, and istherefore, the most widely used material (Da Luz et al., 2016; Oladosu et al., 2016).
Screening of existing landraces as well as creation of new mutant variety withimproved salt tolerant traits is also a major goal for future study. In this study, we
aimed to describe the possibility to get mutants with higher yield, photoperiod
insensitive, and shorter plant height from local T. aman rice genotypes Ashfal and also
to get mutant with higher yield and early maturity from BRRI dhan29 and it is extended
to obtain mutant from NERICA10 in rice by ion beam irradiation.
Materials and Methods
Hulled rice seeds of Ashfal, BRRI dhan29 and NERICA10 were exposed to 26.7
MeV/n carbon ions with different doses of 0, 10, 20, 40, 60, 80, 100, 120,140, 160, 180
and 200 Gy at Japan Atomic Energy Agency (currently QST, Takasaki, Gunma,
Japan). One hundred and fifty seeds were used for each irradiation dose. The
irradiated seeds were allowed to germinate in petri-dishes. Germinated seeds weresown in seed bed to grow seedlings. Seedlings were transplanted in the field. Single
seedlings were transplanted into each hill. Seedlings were transplanted at the
PBGED experimental field, IFRB, AERE, Savar, Dhaka, and also at the experimental
field of BINA, Mymensingh. Plot size was 5.0 x 2.0 m2 and distance between rows andplants were maintained 20 cm and 15 cm respectively. The experiments followed RCB
design with three replications. Recommended dose of fertilizers, cultural andintercultural practices were done as and when required. Porous pipe was depth into
15 cm of soil to maintain irrigation in rice field. All fertilizers were applied during
3

final land preparation excluding urea. Urea was applied at three equal installments
at 7, 30 and 55 days after transplanting. Three hand weeding were made and the plots
were kept saturated with irrigation till maturity during Boro season. Heights were
measured randomly from ten mutant plants of each irradiated dose at the time of
harvesting. Number of days to heading was recorded as the number of days required
from sowing time when 50% of plants of each line headed. Days of maturity wasrecorded as the number of days required from sowing time when 90% of plants of eachline appeared with yellowish grains. These two data were recorded through visualobservation by visiting the plots every alternate day. Number of effective tillers perplant and panicle length was measured at the time of maturity with randomly
selected five competitive plants. Filled and unfilled grains/panicle was recorded fromfive hills per plot at harvest. Grain yield was recorded from 1 m2 area which was later
converted to yield at ton/ha at 14% moisture. Moisture data was recorded with a grainmoisture meter.
Results
Table 1 showed that BINA dhan18 was cultivated at seven different zones in
Bangladesh to observe the growth uniformity and yield performance. Table 2
indicated that data of yield attributing traits of mutant (RM(2)-40(C)-1-1-10) andparent (BRRI dhan29) are non-significant. Maturity period and yield was foundsignificantly different between mutant and parent and also among the location
cultivated in Bangladesh (Table 3). It was observed that highest (7.8 t/ha) yield withshort (145 days) duration was observed at mutant grown at Pakkhifanda, Rangpur
area over the parent. Table 3 also indicated that mutant yielded higher and required
short duration to mature over parent at every location grown in Bangladesh. Table 4
proved that mutant produced long grain fine rice as grain length was recorded 9.0(7.4) mm over parent 8.0 (6.5) mm.

4

Table 1. Multi location trial of BINA dhan18 at different sub-station of BINA.

Location

Date of seed sowing

Age ofDate oftransplanting

seedlings(days)

BINA farm, Mymensingh

02 December 201309 January 201439

Maijbari, Mymensingh

02 December 201310 January 201440

02 December 201314 January 201444

02 December 201314 January 201444

01 January 2014

12 February 201443

01 January 2014

13 February 201444

02 December 201331 January 2014

61

BINA sub-station farm,
MaguraFarmer’s Field, Magura
BINA sub-station farm,
Rangpur
Pakkhifanda, Rangpur
BINA sub-station farm,BarisalTable 2. Yield attributing traits of mutant (RM(2)-40(C)-1-1-10) over parent (BRRI
dhan29).
Mutant/variety

Plant

Effective

PanicleFilledUnfilledheighttiller (no.)

lengthgrains/grains/(cm)

paniclepanicle

(no.)

(no.)

(cm)
RM(2)-40(C)-1-1-10

101

11.42

24.7

113

33

BRRI dhan29

100

11.87

24.5

111

33

LSD(0.05)

NS

NS

NS

NS

NS

NS- not significant

5

Table 3. Comparison of maturity period and yield of mutant (RM(2)-40(C)-1-1-10) withthe parent variety (BRRI dhan29) at different sub-station of BINA.
Location

Maturity period (days)
RM(2)-40(C)Yield (t/ha)

BRRI dhan29-1-1-10

RM(2)-40(C)-1-1-10

BRRI dhan29

BINA farm, Mymensingh

150

165

6.9

6.5

Maijbari, Mymensingh

152

165

5.8

6.3

157

168

7.6

5.7

155

168

5.8

5.5

155

165

7.5

7.1

145

160

7.8

7.2

155

170

5.1

4.9

Farmer’s Field, Barisal

151

162

5.2

5.2

LSD (0.05)--

Average

153

165

BINA sub-station farm,
MaguraFarmer’s Field, Magura
BINA sub-station farm,
Rangpur
Pakkhifanda, Rangpur
BINA sub-station farm,Barisal

0.36
6.46

6.05

Table 4. Comparison of grain characters of the mutant (RM(1)-40- (C)-1-1-10) and theparent variety (BRRI dhan29).
Mutant/variety

1000-grain

Grain length

Grain breadth

Length:weight (g)

(mm)

(mm)

breadth ratio

RM(1)-40(C)-1-1-10

26.0

9.0 (7.4)

3.0 (2.5)

3.0 (2.96)

BRRI dhan29

20.6

8.0 (6.5)

2.5 (2.2)

3.2 (2.95)

Numbers in the parentheses indicate the values of dehusked grain.

6

Control (NERICA10)

BINAdhan19 (Mutant)Discussion
On the basis of lethality, ion beams induced higher frequencies of mutation than
gamma rays. In view of this, seeds of Ashfal rice, BRRI dhan29 and NERICA10 were
irradiated with different doses of carbon ion beams and were grown in Boro and Aman
season. These results showed that photoperiod sensitivity was altered by carbon ion
irradiation in case of BINA dhan14 whilst it was developed from highly photoperiod
sensitive cultivar Ashfal. It was also found that plant heights of the three studied
land races at M2 progenies were significantly shortened than that of their parents.
This is also observed consistent with the fact that the fertile M 2 plants had shorter
plant height than their parents. This might be due to the fact that the fertile mutant
lines shifted from vegetative phase to reproductive phase earlier than parents and it
is caused by ion beam irradiation. It has been reported that one dominant and one
recessive gene are responsible for photoperiod sensitivity in rice (Yu and Yao, 1968;
Yokoo and Fujimaki, 1971; Yokoo et al., 1980). It could be happened that carbon ion
beams irradiation inactivated or down regulated the activity of the dominant gene in
the irradiated seeds which makes the mutant lines homozygous monogenic recessive
for photoperiod insensitivity. BINA dhan18 showed higher yield and early maturity
compared to parent of BRRI dhan29 which could be due to the useful effect of carbon
ion beam. BINA dhan19 also showed higher yield, early maturity and highly droughttolerant compared to parent of NERICA10 which also thought to the useful effect ofcarbon ion beam. It was obvious from the study that BINA dhan14 was developed

7

using 200 Gy and BINA dhan18 and also BINA dhan19 was developed using 40 Gy
respectively through carbon ion beam irradiation. The rest of the doses of theparticular genotypes under investigation were omitted from these evaluations
because they were prone to lodging for weak culm although they had shortened plants.
Moreover, these progenies had shorter panicle with smaller grains and lower numberof seed grains and also their inferior performance. Although the grain characteristics
such as size and shape altered in case of most of the mutants but grain weight almost
similar as the parent. Hence, it could be concluded that it is possible to induce fixedmutants in indica type rice Ashfal, BRRI dhan29 and NERICA10 through 40-200Gy
doses of carbon ion beams irradiation. This result is also in agreement with the resultsof Honda et al. (2006) who reported genetically fixed mutants in M1 generation ofsweet pepper for dwarf height and yellow pericarp. Therefore, all of these suggest thatheavy ion beams irradiation has unique properties that induce fixed mutation.
Acknowledgements
This work was carried out under the Forum for Nuclear Cooperation in Asia
(FNCA) and IAEA/RCA Mutation Breeding Project. Special thanks to Dr. Atsushi
Tanaka, Dr. Hase, National institutes for Quantum and Radiological Science andTechnology (QST); Ms. Aki Koike, Nuclear Safety Research Association (NSRA); Dr.
Liu Luxiang, RCA Coordinator of China for their kind help and cooperation.

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