GRM 2013: Delivering drought tolerance to those who need it: From genetic res...
Mohammed ayaad
1. Molecular Genetic Assessment of some
Rice Mutants Induced by Gamma
Irradiation
المس األرز طفرات لبعض الجزيئي الوراثي التقييمتحدثة
جاما أشعة بواسطة
by
Mohammed Hussein Essa Ayaad
2. Under supervision
Dr. Eman Mahmoud Fahmy
Prof. Emeritus of Genetics, Genetics Dept., Faculty Of Agriculture,
Ain Shams University.
Dr. Nermin Mahmoud Abd El-Gawad
Prof. of Genetics, Genetics Dept., Faculty Of Agriculture, Ain
Shams University.
Dr. Raafat Anwar Kamal Moustafa
Prof. Emeritus of Genetics and plant breeding., Plant Res. Dept.,
NRC, Egyptian Atomic Energy Authority.
3. Rice is the most important food crop in the world.
More than half of World population
are rice consumers.
About 93% increased will occur in
developing countries.
7 billion
people
2014
8 billion
people in
2025
We must
produce 40%
more rice by
2025.
FAO Database 2014
5. Challenges?
Limited cultivated area.
Limited water resources.
Rice water consumption
(Plant growth duration)
(1k rice = ?? M3 of water)
Increased growth population
Diseases
Rice scenario in Egypt
• 55.5 billion
M3 of water/
year
River Nile
• 620.285
hectares (1.5
MF)
Egypt
• Long
• Medium
• Short growth
duration
World
6. Rice blast disease
Evaluation of blast disease resistance
Rice blast, caused by the
pathogenic fungus Magnaporthe
oryza, is one of the important rice
diseases, causing great damage to
rice yield because of its wide
distribution and destructiveness
under favorable conditions (Ou,
1980)
7. Novel variation can be
produced.
Direct mutant varieties (limited
breeding effort required).
The mutations are generally
random and unpredicted
The rice genome (2n=24) is about
420 Mbp (Indica with 32000
gene) to 466 Mbp (Japonica with
46000 gene).
Rice Mutants
(815 from 3200)
Early maturing
High Tillering
Yield
Quality
Semidwarf
Disease resistance
Cold tolerance
Salinity tolerance
Agronomy
8. Yield evaluation of different rice genotypes in two different
locations for two advanced seasons (M6 and M7).
Screening all genotypes for blast disease resistance under natural
and artificial infection conditions.
Evaluation of aroma and the grain quality traits.
Applying SSRs as molecular techniques to identify causative
mutations and develop suitable molecular markers for good yield-
related traits.
Objectives
9.
10. 1. MATERIALS
Five local rice (Oryza sativa, L.) cultivated genotypes; Sakha101, Sakha102,
Sakha103, Giza177 and Egyptian Jasmine; in addition to eight mutant rice genotypes
in M5 generation; Egy22, Egy23, Egy24, Egy202, Egy32, Egy33, Egy34 and Egy316.
1. Plant Materials
Table (1): The origin and pedigree of the used rice cultivars
Cultivars Pedigree Origin
Sakha101 Giza 176/ Milyang 79 Egypt
Sakha102 Gz4098-7-1 / Giza 177 Egypt
Sakha103 Giza 177 / Suweon 349 Egypt
Giza177 Giza 171/Yomji Egypt
Egyptian Jasmine IR 358 Egypt
12. Mutants The parent
Irradiated
dose (Gy)
Growth duration
Important trait as
individual plant
Egy22 Sakha102 200 Gy 130 days High yielding
Egy23 Sakha102 300 Gy 130 days Highly synchronization of
flowering, red plant.
Egy24 Sakha102 400 Gy 135 days High yielding
Egy202 Sakha102 200 Gy 125 days High tillering, red
pericarp rice, long
stature.
Egy32 Sakha103 200 Gy 135 days High yielding
Egy33 Sakha103 300 Gy 135 days High yielding
Egy34 Sakha103 400 Gy 135 days High yielding
Egy316 Sakha103 300 Gy 110 days The highest 1000 grains
weight
Different mutations pedigree
15. 2. Methods
2.1.Yield-relatedtraitsEvaluation
All genotypes were cultivated in
two locations (RRTC, Sakha, Kafr
Elsheikh and Agriculture Research
Center, Gemiza, Gharbia).
The usual procedures for conventional
production of rice were followed.
1. Heading date
2. Plant height (cm)
3. Panicle length (cm)
4. Number of panicles per plant
5. Number of grains per panicle
6. 1000 grain weight (g)
7. Grain yield per one m2 (g)
16. Microsatellites are highly
polymorphic, genome-specific,
abundant and co-dominant,
and they have become
important as genetic markers
in rice breeding programs.
Thirty two SSRs primers were
used in the present study to
discriminate the genetic variation
among the 13 rice genotypes and
to detect highly polymorphic
molecular markers for some
studied traits.
2. Methods
Rice quality
evaluation
Blast disease
evaluation
Microsatellite
Markers
34. 2.3. Evaluation of grain quality
The traitsof physical qualityof grains
Hulling and milling percentage.
Length, width and grain shape
Head Rice Yield (the weight of
whole white rice after milling as
a percentage)
The cooking and eatingcharacteristicsof rice
Amylosecontent(high; ≥ 25%, intermediate;
20-25% , low; 15-20% and waxy is zero)
Gelatinizationtemperature
Aroma
38. 2.2. Screening of blast disease resistance
Dry grains of the 13 genotypes
were infected by six races of
Magnoporthe oryzae in the glass
house for 30 days after the
cultivation process.
All of thirteen genotypes`
grains were cultivated in the
blast nursery, in two
different locations in the first
of July for two seasons.
Natural infection Artificial inoculation
47. b) RM7049
c) RG28
a) RM23120
Marker
Sk101
Sk102
Sk103
Gz177
Egy23
Egy202
Egy316
Egy22
Egy24
Egy32
Egy33
Egy34
E.Jas
2. SSRs analysis for Aroma
Sk103
Egy202
The primer RM23120 showed 4
alleles, followed by the primer
RG28 presented 2 alleles as 202
and 121 bp. While RM7049
revealed one allele (70 bp).
RG28 produced one common
band with molecular size 202 bp
among the used aromatic
genotypes (E. Jas, Egy34, Egy33,
Egy32 and Egy24) except Egy22.
52. Conclusion
The mutant Egy316 was earlier
than other genotypes (110.9 days)
and recorded the highest 1000
grain weight (32.7g).
Egy202, showed the highest
number of panicles/m2 (922.8),
but the grain yield (ton/fed)
decreased as a result of lodging.
Maximum grain yield (ton/fed)
appeared for Egy22.
The studied rice mutants except Egy202
and Egy23 had slender grain shape in
addition to the Egyptian Jasmine (more
than 3 mm), while the cultivars; Sk101,
Sk102, Sk103, Gz177 and Egy23, as well
as Egy202 and Egy23 were medium grain
shape(2.1 to 3.0 mm).
All genotypes have intermediate amylose
content except Egy23, Egy202 and Egy24
(high amylose content).
The strongest aromatic genotype
was Egy22 as compared to the
local aromatic cultivar (E. Jas).
Under natural and artificial infection for
blast disease, the 13 rice genotypes were
resistance and moderate resistance
except Sk101
53. The primer RM125 may be specific primer for
grain shape.
The primer RM135 gave one positive
marker in the high tillering mutant
(Egy202); it may be related to the studied
trait.
The primer RG28 was able to recognize
between aromatic and non aromatic
genotypes except Egy22.
RM224, cosegregated with blast disease,
were absent in all genotypes except
Sk101.
54. Dr. Abd El-Salam Ebaid Draz
Dr. Fatthy M. Abd El-Tawab
Dr. Eman Mahmoud Fahmy
Dr. Nermin Mahmoud Abd El-Gawad
Dr. Sabah M. Hassan
Dr. Raafat Anwar Kamal Moustafa
Dr. Sobieh S.S. Sobieh
Dr. Abd El-Magid M. Shaheen
Dr. Osama El-Badawy
Dr. Rabiea El-Shafey
Dr. Aly El-Gohary
Dr. Yassir El-Refaee
Dr. Aysam Faied
Dr. Kamal Abd El-Lateef
Dr. Nesrin Nazmy
Acknowledgment
To my family,
My father,
My mother,
My wife
My boys (Mohanad and Eiad)
Dr. Khaled Elazab,
Dr. Ibrahim Osamy,
Dr. Shreif Yousif,
Dr. Mohammed Basyouny,
Dr. Mohammed Abdelrazek,
Dr. Waleed Korany
Mr. Mohammed Ali
