Annex 4

Q&A Sheet

FNCA MB Workshop Question & Answer Sheet
Reporter /Question / Comment

Answer / Response

Bangladesh

1) Thank you for the nice presentation! In1) Thanks a lot Mr. Aurigue for your kind(Dr. Mamun)

Slide 7, do the leaves of the mutant lines

observation. In slide 7 you are right due tovary from one another or was it due tolight duration color looks different. Another

difference in lighting during photography? Inon slide 6 left side you can see the control

Slide 9, the color of the grains are distinctly

plants photographs, Mutants are earlier indifferent from one another. Is there nocase of flowering and maturity. Thanks.

Answererpicture of the control or untreated seedlingsin the plot?
(from Mr. Aurigue, PHL)

China1) I am just wondering whether tissue

1) Yes, tissue culture and transformation(Prof. Shu)

culture process in genome editing and/or

could

doubled haploid production may induce

genome edited plants, we had assessed thecertain amount of somaclonal variation. Imutation

hope it is a negligible level, but do you havegenomes of regenerated plants and came toany data on this point?

the conclusion that Agrobacterium-mediated(from Dr. Hase, JPN)

transformation (for genome editing) induced

inducegenomicfrequencybyvariations.

Forsequencing

5

yourmutations at frequencies similar to routine

accomplishments particularly the numerous

tissue culture (Li et al 2016, Zhejiang Univ-publications that are useful in mutation

Scibreeding! I look forward for more relevant

17(12):992-996. For anther culture, we

documentation of your works.

believe the tissue culture process could also

(from Mr. Aurigue, PHL)

generate somaclonal variation, though we2)

Congratulations

onB(Biomed&Biotechnol)

2016

have not quantified this kind of variation.
However, we did observe variations, such as
male sterile plants, in a DH2 population,
which suggest the existence of somaclonalvariation.
2) Many thanks

Indonesia

1) What is the major determinant of high1) High yielding in Kemuning 1 and 2 was(Dr. Sobrizal)

yielding in “KEMUNING 1&2”?

determined by combination of number of pod- No. of seed pod per plant

per plant and grain size, which were higher- Grain size

than their parent and checks. In addition,- Biotic/abiotic tolerance

Kemuning 1 and 2 were developed to1

- or others?

drought stress tolerance so both are suitable(from Dr. Hase, JPN)

for drought prone area.

2) What kind of situation is "shade tolerant"?

2) The study of shade tolerance is carried(from Mr. Takahashi, JPN)

out under 50% of light intensity which is3) Congratulations for developing newmeasured by lux meter.

mutant varieties of soybeans! May I know if3) Thank you! The parent of Kemuning 1 andthere is any drought-tolerant variety used as2 doesn’t exhibit drought-tolerance. We

control or check for Kemuning 1 andused drought-tolerant variety as a check and

Kemuning 2? Does the parent, Pandermen,

the result showed both Kemuning 1 and 2

exhibit drought tolerance? Also, I wanted toshowed better performance than the check.

confirm if Sugentan 1 and Sugentan 2,Wewhich mature very early, are NOT drought-Sugentan 1 and 2, so we cannot confirm

tolerant. Thank you for your replies!

about that.

didn’tdodrought-stress

testfor(from Mr. Aurigue, PHL)

Japan1) This is my first experience to join FNCA1) Thank you for your interest to use ion(Dr. Hase)

meeting and I am very happy to have thisbeams. You can do so if the purpose is in line

opportunity and know about the progress ofwith the FNCA project and also the PL (Dr.

several Asian countries in the use of nuclearSobrizal) agrees to use. Probably, I canin plant breeding.

provide a beam time for the FNCA project inI am interested to join in the use of ion

April or May 2021. Details will be announced

beams for mutation breeding, especially forin February. Please note that the beam timesoybean mutation, which was mentioned byis limited, and therefore, I may not be

Dr. Hase. How can I include genetic material

possible to irradiate all the seeds. You needin this program?

to obtain phytosanitary certificate from the(from Dr. Puspitasari, IDN)

competent authorities of your country to2) Dr. Hase, have you published any of theexport soybean seeds to JPN. I havereports that you presented in our previous

irradiated soybean seeds from Vietnam inFNCA Meeting Workshop? I will appreciate

the last two years. Information from Dr. Thao

it very much if you could provide us thewill be helpful to determine proper dose.

details of any publication or the link for an e-2) Thank you for your interest in our work,

copy.

Mr. Aurigue. Here is the link for my recent(from Mr. Aurigue, PHL)

publications that I talked at the workshop.
https://www.frontiersin.org/articles/10.3389/f
pls.2020.00336/full
https://www.nature.com/articles/s41598018-19278-1
https://academic.oup.com/jrr/article/61/5/632

9/5881841

Korea

1) In one of your slides it was mentioned

1) A new variety with high function means(Dr. Kang)that developing a new variety with highthatfunction such as rice, can you explain whatcompounds,

is meant by high function on what charactercompounds varies depending on the crop.

/ trait you want to develop?

Main functional compounds in recently

2) For the commercialization of this new

developed rice were tocopherol, amino acids

mutated variety, what approach do you use

and pigments, etc.

so that it can be accepted quickly by farmers

2) New mutant varieties of rice andand the community?

soybeans, which are officially registered, areitcontains
andalot

theoftypefunctionaloftarget

andusually propagated by ourselves and thenmetabolomics study, in what generation ofdistributed directly to farmers. Other crop

your mutant population was this approach /seeds and technology developed by ourstudy carried out?

research team have been transferred to the(1)-3) from Dr. Dewi, IDN)

industry sector.

4) Do you have any plans to use phenomics

3)

in mutant screening? I think it is a biggenomics and metabolomics analysis tochallenge but if it works, we could find new

selected mutants or population in M2~M6

mutants that cannot be identified bygeneration.

conventional methods.

4) I think that the KAERI research team will(from Dr. Hase, JPN)

be working on setting the conditions for the5) Dr. Kang, do you have any publication

meantime. I hope to see much progress inabout the mutant varieties of ornamental

this research in the future.

plants? They are all very interesting and I

5) We published some papers aboutwould like to know more about the details.

mutationThank you!

horticultural plants already. If you search my(from Mr. Aurigue, PHL)

name or keyword, you can easily find it.

Malaysia

1) Do you have a mutation breeding

1) Yes we have. Basically, the mutation(Dr. Hussein)program to improve local rice varieties inbreeding

Malaysia to support food self-sufficiency ingenerated when local verification trials (LVT)

the region, given that local rice has specificare conducted in collaboration with thecharacters, especially taste and aroma,industry players. Once rice mutant shows

which is maintained and developed bysignificant increase in yield, the farmers

farmers in the area?

would request to be part of the mutation

2) Are there any difficulties in propagating

breeding project. Subsequently, the best

different types of mutant varieties so thatmutant variety is maintained and developed

they can be distributed to farmers?

by the industry players.

3)

For

functional

genomics

3We

havebeenbreedingmainlyin

programconducted

ornamental

isorautomatically(1)-2) from Dr. Dewi, IDN)

2) Propagating different type of mutant is not3) Sorry to ask about such minor details, butthe major problem because it can be done

what the graph on the right side in your slide

through research collaboration with the#13 represents? In that, NMR152 andindustry players. The major problem is theNMR122 show the value of 134 and 122,

requirement to seek for approval from therespectively.

national technical committee before the rice(from Dr. Hase, JPN)

variety is allowed to be distributed to the4) Nuklear Malaysia truly deserves thefarmers. Moreover, the approval process isFNCA

tedious and time consuming.

Awardbecauseoftheaccomplishments that have high impact to3) The graph shows in slide no 13 is the SSR

farmers and to Malaysian economy. At thefragment analysis to differentiate between

same time, you are able to promote

the parent and NMR152. 134 refers to themutation breeding technology through yourSSR analysis for NMR152 while 122 refers

publications, Farmers’ Field Day, and press

to the SSR analysis on the parent (MR219)

releases. Congratulations! What is also very

control.

good about the mutants is that it helps

4) First and foremost, thank you to Mr.

protect the environment and the health ofAurigue for the compliments and showing

people due to minimal use of chemicals

great appreciation towards the output ofagainst pests and diseases.

Nuclear Malaysia mutation breeding project.

(from Mr. Aurigue, PHL)

Nuclear Malaysia would also like to express
utmost gratitude to FNCA for the greatsupport.

Mongolia

1) In Slide 12, the mutant lines of wheat

1) Thank you for question. Standard wheat(Dr. Noov)were classified as early maturity (81 and 83

maturity groups in Mongolia are: earlydays), medium maturity (86 and 87 days),maturity 80-84 days, medium maturity 85-90

and late maturity (92 days). Please clarify

days, and late maturity over 91 days. The

your mutation breeding objective stated inslide 4 illustrates our overall objective toSlide 4 that short vegetation period is 80 toreduce maturity days in all maturity groups.

100 days. Why is 92 days considered

For example early maturity not more than 80

already late maturing when it still falls underdays, late maturing lines less than 100 days.

short vegetation period? Thank you!

But, we understand that expression in slide(from Mr. Aurigue, PHL)

4 was not clear to explain above objective.

The1) Based on your framework, what are your

1) Selection criteria is acceptable phenotype

Philippines

considerations for screening abiotic stress(good plant architecture, stature, early(Mr. Cabusora)

in the M4 generation?

maturing, exserted panicle, etc.) at the M2(from Dr. Dewi, IDN)

population. Since our wildtypes are abiotic

2) Do you see any difference between thetolerant landraces with poor phenotypes.

4mutant population generated by seedThe advancement to two more generations

irradiation(up to M4) is done only to evaluate theandinvitro

mutagenesis,including unfavorable phenotype?

uniformity of the lines and the stability of the(from Dr. Hase, JPN)

mutation. We need to be sure that themutation is stable (genetic) and that the lines
will not revert back to its original phenotype(epigenetic) on the next generations. So
basically the screening for stresses, is toselect mutants with good phenotype andretained abiotic stress tolerance.
2) Actually no, Sir. We don’t observe any
difference in terms of variability induction.
The only advantage of using IVM is that at asmall population, variability is high and wewere able to select best lines (higherefficiency). Unlike in seed mutation we need
thousands of M2, only to have small
selection efficiency.
Thank you for the questions, Dr. Hase. If
may I request your email address. I would
like to consult some matters regarding oneof our mutant exhibiting novel floral mutationdifferent from those already published.
Thank you.

Thailand

1) Thank you for joining the workshop. It

1) Hello Dr. Hase. Yes, of course. Next time,(Mr. Noenplab)

was nice to meet you and I am looking

we will talk about flood tolerant and drought

forward to hear your research work more intolerant elite lines developed from mutation

detail next time.

breeding in our pipeline. Thank you for(from Dr. Hase, JPN)

hosting the event.

Vietnam

1) How to use brown and black soybean?

1) Thank you for the question. In Vietnam(Dr. Le)(from Mr. Takahashi, JPN)

previously only yellow soybeans were used

2) The results of gamma irradiation on

for food. Black soybean DT215 is the first

peanut are very good and interesting.

black soybean variety created and planted in

However, could you please ascertain which

Vietnam. DT215 has high levels of omega-3,chlorophyll mutation was observed? Fromomega-6,

the pictures presented in the slides, it does

processing functional foods. Currently, thisnot look like an albina (white) at all. You may

variety has been transfer breeder copyright5andcarotene,

suitableforwant to check if it is actually xantha or

to a food company, which processes bottled

chlorina because albina hardly survives due

food drinks and soybean cakes rich into lack of chlorophyll. Thank you!

nutritious dini.

(from Mr. Aurigue, PHL)

2)3-1) Responding to Dr. Hase's answer to my

implementation process, not only peanuts

question, I would like to ask about thebut soybeans are also often obtained albino

proper dosage for irradiating soybeansin the M1 and M2 generations. The albino

using ion beam.

variant can be whole body or partial tree;(from Dr. Puspitasari, IDN)

The timing of appearance can be in the3-2) Could you share the data on theseedling or when the plant is mature. Sosurvival rate of soybean seeds irradiated onsome plants still grow and collect seeds.

May 14th, 2019? On that day, I irradiated

These variations are less in M2 and by M3

soybean seeds with the following doses.

almost disappeared. In the presentation, we

320 MeV Carbon: 10, 20, 30, 40, 50 Gy

only give examples of the occurrence of

107 MeV Helium: 20, 40, 60, 80, 100 Gy

mutants without going into further research(from Dr. Hase, JPN)on this mutant type, the main direction for3-3) Dr. Thao, thank you for your response.

screening in our mutants peanut breeding isClear dose-response is seen on the survival

to choose varieties with high lipit content.

rate of M1 seeds.

3-1) It is true that we are having trouble

Dr. Puspitasari, I think this data is helpful todetermining the dose of ion beam irradiation

check the radiation sensitivity of youron soybeans. We are currently evaluating

materials. Let's think about it again, after

the M3 generation, but the seeds numbers isbecoming clear that I can hold a beam timevery small. It seems that we determined thefor this project.

dose too high and the wide distance. So we(from Dr. Hase, JPN)

are working to determine the dose closer

3-4) Dr. Thao and Dr. Hase, thank you very

together in next irradiate to determine themuch for your cooperation. This is very

optimal dose.

useful information. We look forward to apply3-2) Please see table 1 is the data for ion

it to our genetic materials.

beam on soybean. Thank you for your(from Dr. Puspitasari, IDN)

cooperation.

6

Thankyouforquestion.

IntheTable 1
The survival rate of soybean seeds irradiated (%)
1. 320 MeV-C842008-1

842008-36

F3507-2

F3507-3/1

M1M2M1M2M1M2M1

M210 Gy

6187,55786,56383,36687,120 Gy

4886,94588,14586,84685,930 Gy

3088,02987,73187,42786,340 Gy

1787,81786,91987,12085,750 Gy

1087,5987,5987,01187,10 Gy

88,187,287,586,4

2. 107 MeV-He
842008-1

842008-36

F3507-2

F3507-3/1

M1M2

M1M2

M1M2M1

M220 Gy

6788,16985,16788,46685,340 Gy

6087,25787,35887,958

86,060 Gy

4789,148

86,44688,14885,780 Gy

3186,330

87,53387,74187,1100 Gy

2087,42288,6

1987,32186,10 Gy

87,886,3

789,185,6