Nghiên cứu chọn tạo dòng bố mẹ chống chịu bạc lá, rầy nâu phục vụ phát triển lúa lai hai dòng ở việt nam tt tiếng anh

MINISTRY OF AGRICULTURE
AND RURAL DEVELOPMENT

MINISTRY OF EDUCATION
AND TRAINING

VIETNAM ACADEMY OF AGRICUNTURAL SCIENCES
------------------------------------------------------

LE HUNG PHONG

RESEARCH ON BREEDING PARENT LINES TO RESIST
TO BROWN PLANTHOPPER AND BACTERIAL LEAF
BLIGHT FOR DEVELOPING TWO-LINE HYBRID RICE IN
VIETNAM

Major: Genetics and Plant Breeding;
Code: 9 620111

SUMMARY OF DOCTORAL THESIS

HA NOI - 2018


The thesis is completed at:
VIETNAM ACADEMY OF AGRICULTURAL SCIENCES
(VAAS)

Science instructor:
1. Associate Professor. Dr. Nguyen Tri Hoan
2. Doctor. Nguyen Nhu Hai

Reviewer 1: Associate Professor. Dr. NGUYEN THI TRAM
Reviewer 2: Associate Professor. Dr. NGUYEN THI NGOC HUE
Reviewer 3: Associate Professor. Dr. NGUYEN VAN VIET

The thesis is reported at the grassroots thesis dissertation
council meeting at:
Vietnam Academy of Agricultural Sciences (VAAS)

The Thesis can be found at the library:
1. Vietnam National Library;
2. Library of Vietnam Academy of Agricultural Sciences;
3. Library of Field Crop Research Institute25-30.
1.

Vũ V ĐÃ CÔNG BỐ LIÊN QUAN


1
INTRODUCTION
1. The urgency of the topic
In recent years, hybrid rice have contributed significantly to the food
security, making an important contribution to stabilizing production and socioeconomic development of our country, while the world economy has many
changes. However, the expansion of hybrid rice area faces many difficulties
such as imported seeds with high cost and not being able to be active in seed
sources and the set of selected varieties in the country are still few, there are
not many varieties resistant to pests and diseases (resistance to brown plant
hopper, bacterial leaf blight...) and unfavorable conditions for production. The
source of pest and disease-resistant parent materials for tolerance hybrid rice
breeding is poor, and production still faces many difficulties and risks due to

weather, pests. Besides, the area of rice cultivation in our country has been
significantly reduced due to the effects of climate change causing drought and
saline intrusion. The development of pests and diseases (such as bacterial
blight, brown planthopper, white back planthoper, Rice ragged stunt virus, Rice
grassy stunt virus, Southern rice black-streaked dwarf virus .etc.) causing
difficulties for forecasting work and causing great damage to rice production.
The report of the Plant Protection Department (MARD) in 2013,
2014, 2015, 2016 and 2017 shows that: Recently, rice bacterial blight, stripe
and brown planthopper, white back planthopper Rice grassy stunt virus,
Southern rice black-streaked dwarf virus appear and cause serious damage in
both Winter-Spring and Summer / Summer-Autumn crops in the Northern
provinces. The results of the study "Research on breeding parent lines to resist
brown planthoper and bacterial blight to develop two-line hybrid rice in
Vietnam" will create parent lines with good resistance and high combining
ability. To diversify sources of materials to choose to create 2-line hybrid rice
varieties to resist brown planthopper and bacterial leaf blight.
2. The objective of the project
- Selected 1-2 lines of parents capable of resisting tolerance of
bacterial leaf blight, 1-2 parental lines that are resistant to brown planthopper


2
for the selection and creation of two lines of hybrid rice to resist brown
planthopper and bacterial blight.
- Selected 1-2 new, high-yield hybrid combination F1 (7-8 tons / ha),
good quality, resistant to bacterial blight, brown planthoppers for hybrid rice
development in Vietnam.
3. The scientific and practical significance of the topic
3.1. Scientific significance
- The research results of the thesis supplement the scientific basis and

the method to select the creation of parents maternal lines of thermosensitive
genic male sterile (TGMS) and breedingtwo lines hybrid rice with resistance to
brown planthopper and bacterial blight in Vietnamconditions.
- Information on new lines of parental lines to brown planthoppers,
bacterial blight risistance to contribute to the selection of breeds in using
materials to breed and select hybrid rice two lines with resistance to blight and
brown planthopper.
3.2. Practical significance
- Determining the source of materials for the hybridization of the
father and mother line of two lines of hybrid rice resistant to bacterial leaf
blight and brown planthopper.
- The lines of TGMS as AMS35S-KBL, AMS30S-KBL (resistance to
bacterial leaf blight ), KR95S, KR142S (brown planthopper resistance), the
male lines as RP8, RP088, RP3, R1028-KR, R116R (BPH resistance) have
many agronomic characteristics learning, good sexual characteristics, high
combined ability of yield, is a good material source to breeding hybrid rice
varieties resistant to bacterial leaf blight and brown planthopper.
- F1 Hybrid rice two lines combination with resistance to blight,
brown planthopper have a short growth duration, high yield, contributing to
testing and expanding hybrid rice production in Vietnam.
4. New contributions of the topic.
- Successfully create 02 lines of TGMS (AMS35S-KBL, AMS30SKBL) with bacterial blight resistance gene, stable sterility, high rate of stigma


3
exertion, short growth duration, high value of general combining ability about
real productivity.
- Create 2 lines of TGMS (KR95S, KR142S) and 05 lines (RP8,
RP088, RP3, R1028-KR, R116R). These parental lines have a short duration,
good agronomic characteristics, high combined yield ability with brown plant

hopper resistance genes .These parental lines are very good materials for
selectin two line hybrid rice varieties tolerant to brown planthoppers in
Vietnam.
- Selection of 3 hybrid rice combinations of TGMS lines with
bacterial leaf blight resistance genes, having higher yield than check varieties
10% is AMS35S-KBL / R100, AMS30S-KBL / R116 and AMS34S-KBL /
SR18, resistance to blight score 3-5. Throught transferring the bacterial leaf
blight resistance genes into the TGMS lines initially improved the resistance to
blight disease of two-line hybrid rice HYT124 and HYT116.
- Selecting 06 hybrid rice combinations of KR142S / SR3, KR142S /
SR14, KR142S / SR18, KR95S / SR14, KR95S / AIQ6, KR95S / SR3 with
short duration, high productivity (8.6-9.3 tons / ha) and resistance to brown
planthopper.
5. Subjects and scope of research
5.1. Research subjects
- Existing hybrid rice lines and parents of the Field Crop Research
Institute which the author directly and participated in creating, good parent
lines collected in the country and imported.
- The papolations of BC3 - BC6 from hybrid combinations for
selecting parent lines in the direction of resistance to bacterial leaf blight and
brown planthopper.
- Bactrial leaf blight resistant materials were identified from the
International Rice Research Institute (IRRI) such as IRBB5, IRBB7, IRBB21,
IRBB4, IRBB57, IRBB60, IRBB61, IRBB62, IRBB63, IRBB64 ...
- Brown planthopper-resistant materials have been identified: The
brown planthopper resistance lines of the Agricultural Genetics Institute are:
IS1-2, IS1-3, E2-3, E3… and imported materials internal from IRRI like:


4

Mudgo; ASD7; RathuHeenati; ARC 10550; Swarnalata; Chinsaba; T12;
Pokkali; Ptb33; TN1 (infected).
5.2. Location and time of study
In Hanoi and the Northern Delta provinces; 4/ 2012 to 4/ 2018
5.3. Limited scope of the topic
- Select to create parental lines carrying resistance gene for bacterial
leaf blight and brown plant hopper through hybridization and selection from
intermediate materials.
- Select and purify elite lines with many characteristics of a good
parent line to serve breeding.
- Evaluate the ability to combine the traits of selected lines and
parents to serve hybridization, select 2-line hybrid rice varieties to resist brown
leaf and planthopper.
- Test crossing to find new hybrids.
6. Structure of the thesis.
The main thesis has 132 pages in A4 size with 44 tables of data and 7
figures. The thesis consists of 5 parts; Opening (6 pages), Chapter 1: Overview
of research materials (40 pages). Chapter II. Materials, content and research
methods (17 pages). Chapter 3: Research and discussion results (67 pages),
Conclusions and recommendations (2 pages).
CHAPTER I
OVERVIEW OF RESEARCH MATERIALS
The thesis has consulted and reviewed 29 Vietnamese documents and
81 English documents. Related contents include: 1.1. Scientific basis of the
topic; 1.1.1. Hybrid advantages, genetic bases and hybrid superiority in rice;
1.1.2. Male sterility system in selecting two-line hybrid rice varieties; 1.2.
Practical basis of the topic: 1.2.1. The situation of R&D of hybrid rice in the
world; 1.2.2. Research on combining ability; 1.2.3. Situation of research to
create hybrid rice seeds in the country. With the collected data shows:
* Heterosis: A phenomenon in which F1 hybrids originate from

different parents, demonstrating superiority over their parents in vitality,
productivity, quantity such as panicle size, number of seeds per panicle,


5
number of effective panicle ... Hybrid advantage is shown only in the first
generation (F1 generation), (Nguyen Cong Tan et al, 2002). In rice,hybrids
between two indica and japonica subspecies exhibit maximum heterosis and
are evaluated in descending order: Indica x japonica>indica x javanica>
japonica x javanica>indica x indica> japonica x japonica>javanica x javanica
(Yuan LP, 2006).
* Heterosis in rice: Rice yield in rice expressed on vegetative growth
and reproductive growth as root system (Lin and Yuan, 1980); in traits like, the
ability to branch Lin and Yuan, 1980; Chang et al., 1971, duration (Nguyen Thi
Tram and et al., 1994, Xu and Wang (1980), plant height (Singh SP and Singh
HG, 1978), yield and functional components (Virmani et al., 1982) In
morphological and biochemical traits (Chao, 1972; Deng, 1988), physiological
traits such as photosynthetic intensity, respiratory intensity, leaf area
(Murayama et al., 1984, NguyễnThịTrâm (1994), ... TUTL can be expressed at
all stages of plant growth and development (Akita et al., 1986), in deep
resistance diseases (Lin and Yuan, 1980) and unfavorable conditions
(Senadhira and Virmani, 1987).
* The situation of hybrid rice development in China: China was the
first country to succeed in hybrid rice research in 1973, the process of
developing hybrid rice in China divided into 4 main stages: stage 1 (19641975) and the second phase (1976-1990) was the research period to choose to
create sterile lines and complete the 3-line hybrid rice system. Phase 3 (19902000), is the period of strategic development, initiation of super hybrid rice;
Phase 4 (2013-2020), the new development period, the development period of
super hybrid rice reaches 16-19 tons / ha. According to Yuan L.P. (2017), the
average yield of super hybrid rice reached 13.9 - 15.4 tons / ha in Longhui
district in Hunan province in 2013 and Xupu district in Hunan province in

2014. From 1975 to present, there are more than 5000 Hybrid rice varieties are
recognized and expand the cultivated area of over 500 million hectares, which
is an important factor to ensure food security (Zeng et al., 2018).
* Research on creating hybrid rice varieties resistant to Bacterial
bacterial leaf blight disease: In order to improve the bacterial leaf blight


6
resistance of hybrid rice varieties, many resistance genes (dominant resistance
genes) such as Xa25, Xa21, Xa7, Xa4, Xa23 ... have been used to select the
lines of parents resistant to blight. By the backcross and selective method with
the help of molecular markers, Chinese scientists have chosen to create many
lines of hybrid rice parents with high resistance to blight.
In 2000, Chen et al. (2000) successfully transferred the Xa21 gene
into the Minghui63 restorer line. Deng et al. (2006) successfully transferred
two Xa21 and Xa4 genes into the Mianhui 725 restorer line. He et al. (2007)
succeeded in bringing together the two resistance genes Xa21, Xa7 into the
Minghui63 restorer line. The Philippine Rice Research Institute has
successfully switched to MAS genes Xa21, Xa4, and Xa7 into two-line hybrid
rice TGMS1 (PhilRiceGenbank Acc. No. PRT-1), (Chen et al., 2009). Dindo
and colleagues succeeded in transferring the genes Xa7 and Xa21 into the
hybrid parents of Mestizo1 and Mestiza 3 to create pyramided genes including
Xa4 / Xa7; Xa4 / Xa21; Xa4 / Xa7 / Xa21 at maintenance lines and restorer
lines of Mestizo1 and Mestiza3, Dindo et al. (two thousand and thirteen). Ji and
the team transferred the Xa23 resistance gene from the CBB23 strain into the
restorer line HN189 with the rice blast Pi1 gene, then selected two new restorer
lines, HBH145 and HBH146 with higher resistance to blight. HN189, Jiet. al.
(2014). Luo and his colleagues successfully transferred 2 genes of Xa4 and
Xa21 blight resistance to Mianhui 725 (MH725) line which obtained Wanhui
421 (WH421) line with 96.9% genome compared to MH725 but high leaf

resistance than the original line, Luo et al (2016). Ramalingam et al (2017),
successfully transferred three genes (xa5, xa13 and Xa21) into the maintenance
stream (CO 2B, CO 23B, and CO 24 B). Kumar and colleagues improved the
resistance of the RPHR-1005 strain when transferring 2 blight resistance genes
Xa21, Xa33 and Pi2 from the DRRH-3 strain to the RPHR-1005 line and
selected 10 lines in the ICF4 genome with all 3 The above gene has a good
resilience, Kumar et al. (2016).
Ji et al. (2016), has gathered three rice blast resistance, bacterial leaf
blight and BPH resistance genes into the three-line hybrid rice restorer line,
and the HR13 strain carried all three resistance genes and four lines HR39,


7
HR41, HR42 and HR43. The gene is resistant to blast and blight but there is no
gene for brown planthopper resistance. The Zhongzu 14 series is the result of
all three rice blast resistance genes, bacterial leaf blight and brown plant
hopper. Xiao et al. (2016), transferred a Xa23 gene and two genes for BPH
resistance (Bph14 and Bph15) into the Huazhan restorer line to improve the
resistance to bacterial leaf blight and BPH of Tianyouhuazhan variety. (Ni et
al., 2015), successfully improved the resistance to blast and bacterial leaf blight
of P / TGMS Guangzhan63S series (GZ63S) but still retained the
characteristics of the original maternal lines and F1 hybrids when collected.
Blast resistance gene Pi9 and Xa23 blight resistance gene into GZ63S.
* Researching to create hybrid rice varieties resistant to brown
planthopper: Liet al's report (2011), showed that the team succeeded in
transferring two Bph14 and Bph15 resistance genes into the restorer line of
Minghui 63 (MH 63) and obtained two lines MH63-15 and MH63-14 & 15. H.
Ji et al. (2013) published the results of cloning of BPH resistance gene Bph18.
Ling and Weilin (2016) suggest that the genes Bph14, Bph26, Bph3
and Bph29 were evaluated as the best pest resistance gene. According to Xiao

et al. (2016), BPH is the heaviest rice pest in Asia. The authors transferred 13
genes and QTLs (Bph14, QBph3, QBph4, Bph17, Bph15, Bph20, Bph24, Bph6,
Bph3, Bph9, Bph10, Bph18 and Bph21) into rice lines 9311 by back crossing
with support of the instructions molecule. All lines containing resistance genes
reduced the development of brown backed rice plant hoppers at the plating
stage. Wang et al. (2016) collected two Bph14 and Bph15 resistance genes into
one Huahui 938 restorer line to improve resistance to brown plant hoppers.
Wang et al. (2017) gathered Bph6 and Bph9 genes into line 9311, resulting in
the creation of hybrid rice LuoYang69 with the progeny of 9311 carrying two
highly resistant Bph6 and Bph9 genes with brown backed rice plant hoppers
and unchanged from the original. According to Fan et al. (2017), chose to
successfully create a hybrid line for hybrid rice with the large panical
geneGn8.1, Bph6 and Bph9 BPH resistance genes and Rf3, Rf4, Rf5 and Rf6
recovery genes thanks to molecular markers.


8
* Research results and production of hybrid rice in Vietnam:
According to the Report of the Department of Crop Production, Hybrid rice has
so far been cultivated in most rice growing areas of our country, in which the
northern provinces occupy a large area and mostly. However, in recent years,
hybrid rice production in the Northern provinces has continuously decreased in
both area and yield, reducing both Spring and Summer crops, but the main
cause is determined by the supply of hybrid seeds. processing, high price of
quality hybrid rice seeds, hybrid seed yield produced in the summer season is
low because of pest infestation. At present, we have mastered the technical
process of F1 seed production and multiplication of the female lines of most of
3 lines hybrids based on BoA and II32A –CMS lines. The two line hybrids
developed in Vietnam has been recognized by MARD. In the period of 2005 2009, the production of hybrid rice varieties tended to decrease. However, from
2015 up to now, the F1 seed production area has increased again, each year

area of F1 seed production have reached more than 2200 ha with hybrid seed
productivity 2.1 - 2.7 tons / ha (Department of Crop Production, 2016).
However, the production of hybrid rice seeds in the Northern provinces faces
many difficulties due to negative impacts of climate change, epidemics (black
stripe dwarf in 2017, many areas are lost in Nam Dinh and some Northern
province) so productivity is not high. Production in the South Central Coast
and Central Highlands provinces is more productive but the cost of seed is high
and transportation costs to the North are also high, Large enterprises are not
keen to invest in domestic production, these are The main cause of limited
development of F1 seed production in our country today.
* Research results on pest resistance: After testing the standard lines
carrying the blight resistance gene of IRRI with 11 bacterial lines in the
Mekong Delta Rice Institute, the Vietnam Institute of Agricultural Science and
the Hanoi Agricultural University No. 1 showed the lines IRBB4 (Xa4), IRBB5
(xa5), IRBB7 (Xa7) and IRBB21 (Xa21) are well resistant to bacterial blight
pathogens (6-9 lines). Breeding program to select breeders using molecular
Marker to transfer the Xa4, xa5, Xa7, Xa21 genes into the hybrid rice lines of
the above research units. Vu Thi Thu Hien et al. (2007), transferred the Xa21


9
gene into the TGMS 103S maternal line by hybridization and MAS help to
create hybrids resistant to blight of Viet Lai rice variety 20. Nguyen Van Hoan
(2005) Select breeding Viet Lai 24 hybrid rice with Xa21 gene resistant to
blight disease. The above achievements are the scientific and practical basis for
application in the research and development of two-line hybrid rice in
particular and hybrid rice in Vietnam.
CHAPTER II
MATERIALS, CONTENTS AND METHODS OF RESEARCH
2.1. Research materials

* Materials for selecting parent lines to resist bacterialblight (BB):
- The material lines imported from the International Rice Research
Institute (IRRI) with BB risistancegene such as IRBB4, IRBB5, IRBB7,
IRBB21, IRBB 60,
- 10 BB-risistant populations were backcrossed in BC3-BC5 among
TGMS lines with IRBB4, IRBB5, IRBB7 and IRBB21 lines.
* Materials for selecting parent lines to resist brown planthopper:
- Material lines with BPH resistance gene were identified from
research units such as: Vietnamese Agricultural Genetics Institute (IS1-2, IS23, E2-3, E-1, E-2, E-3, E-4, E-5, E-6, E-7, E-8); Source of imported materials
from IRRI (Mudgo, ASD7, Rathuheenati, ARC 10550, Swarnalata, Chinsaba,
T12, Pokkali, Ptb33, TN1- Contamination control)
- 5 Brown planthopper-resistant populations in the BC2-BC3
generation of the combinations are the lines of IS1-2, IS2-3, E-1, E-2, E-3 ...
carrying the BPH resistance genes with the TGMS and male. lines
* Materials for hybrid rice resistant to bacterial blight, brown planthopper
resistant breeding:
- The TGMS lines such as: AMS30S (827S), AMS34S, AMS35S,
D116S, D67S, D64S, D52S, D59S, TGMS lines checks are T1S-96.
- New lines of parents resistant to bacterial blight and brown
planthopper have been selected.
- The father lines in the source nursery of the hybrid rice R&D center,
the pure rice varieties are resistant to BLB and brown planthopper.


10
- Check variety: TH3-3 (Two – lines hybrid rice)
2.2. Research content
2.3.1. Breeding to parent lines containing the BPH resistance genes
2.3.2. Breeding to parent lines containing the BLB resistance genes
2.3.3. Breeding new hybrid rice towards resistance to bacterial leaf blight and

brown planthopper
2.3. Research Methods
2.3.1. Evaluate the source of materials resistant to bacterial leaf blight
and brown planthopper
- Evaluate some agronomic characteristics of research materials;
- Evaluation of artificial resistance to bacterial leaf blight and brown
planthopper of the research material carrying resistance gene according to IRRI
method 1996, 1997, 2002
2.3.2. Breeding of parental lines resistant to bacterial blight, brown
planthopper
2.3.2.1. Selective breeding of parental lines resistant to brown plant hopper and
bacterial leaf blight :
2.3.2.2. Check the presence of resistance gene in parental lines resistant to
brown plant hopper and bacterial leaf blight
2.3.2.3. Evaluate the combined ability of parental lines resistant to brown
backed rice plant hopper and bacterial leaf blight
2.3.3. Breeding hybrid rice combinations towards resistance to bacterial
leaf blight and brown planthopper
- Using TGMS lines and male lines with at least one parent resistant
to bacterial leaf blight or/ and brown plant hopper to do test cross with other
parents and create two line hybrids resistant to brown plant hopper or/ and
bacterial leaf blight .
- Doing 2-3 sowing dates of parental seeds, transplanting and taking
care normally. When the male and female plants are heading, the parent plants
are uprooted and planted 1 male plant and 1-2 mother plants in 1 pot, when
both male and female plants flowering, do shaking the pollen parents by hand
4-5 days continuously to do pollinate for female parents


11

- Evaluation of agronomic characteristics, yield of F1 hybrids by IRRI
method, 1997.
- Yield comparison experiments are applied according to the National
Technical Regulation on cultivation value testing and use of QCVN 01-55:
2011 / BNNPTNT of Ministry of Agriculture and Rural Development.
2.3.6. Methods of data analysis
Kempthorne model analysis capabilities (1957) and Program analysis
of variance "Line * Tester" Ver.2.0 and Ver.30 software by Nguyen Dinh Hien
(1995, 1996).
2.3.7. Methods and criteria for evaluating pests and diseases
Evaluation of agronomic, pest and yield characteristics according to
Standard Evaluation System for Rice of the International Rice Research
Institute (IRRI, 1996).
CHAPTER III
RESEARCH RESULTS AND DISCUSSION
3.1. Results of breeding and selection of parental lines resistant to brown
planthopper
3.1.1. Results of evaluating materials resistant to brown planthopper:
After evaluating the resource of BLB resistant genes and review the
published documents in Vietnam and IRRI we chose CR203, IS1.2, E2, E3,
E6, E7, RathuHeenati, Swarnalata, Pokkali as materials for for breeding
parental lines resistant to BPH .
3.1.2. Breeding, selecting parental lines resistant to brown planthopper
The selection process of BPH resistant male lines is carried out
according to Figure 2.1 (page 53). Through evaluation, selection and
purification until 2014, there are 9 most promising lines viz. RP088-51,
RP088-148, RP088-48, R253-KR, R1028- KR, RTQ5-KR, R838-KR, RP3,
RP8 having many good characteristics of male lines, were selected for further
and more specific studies:
The results of evaluation of resistance to brown planthoper throught

artificial infection, the resistance grades of brown planthopper infestation in
2017 at the Plant Protection Institute showed that: RP8 line is resistant to


12
brown planthopper (score 3), other lines with resistance genes are: RP088-148,
RP088-48, RP3, R1028-KR med- resistance (score 5).
Table 3.7. Results of evaluation of resistance grates to BPH
of some parent lines in 2017
No

Name

Cross source

Score

Resistant
grade

1
2
3
4
5
6
7

D116ST/E3-4-1
D52S/Pok6-427

D59S/E3
TL6/E6-148-1
D116Str/E3
AP9
P20-5
Minhhui
63/IS1.2-8
R9311/IS1.2-3
R1028/E3-60-2
TL6/E7-48-3
AMS30S/RP3
AMS30S/R116
-

5
5
5
5
5
7
7

MR
MR
MR
MR
MR
S
S

3

R

9
10
11
12
13
14

KR95S
KR142S
D59S-KR
RP088-148
D116tr-KR
AP9
P20-5
RP8
(APIRX14)
RP3
R1028-KR
RP088-48
HYT285*
HYT116**
TN1

5
5
3

5
3
9

MR
MR
R
MR
R
SS

15

Ptb33

-

3

R

8

Note
Female
Female
Female
Male
Female
Male

Male
Male
Male
Male
Male
Hybrid
Hybrid
Check (S)
Check (R)

Source: Institute of Plant Protection, 2017; Note: * AMS30S / RP3
combination has male line (RP3) with BPH resistance gene; ** HYT116 is
two-lines hybrid rice has male line of resistance BPH, R116
* Assessment of general combining ability (GCA) of new TGMS
Table 3.9. The general combine ability (GCA)

Testers
(TGMS)

Lines
(Male

Of some male lines
Testers/ lines
CT1-AMS35S
CT2- AMS30S
LSD0,01
LSD0,05
TH29
TH12

GCA
-0,547
0,547
8,462
6,234
0,642
-1,425


13
lines)

M415
M359
TH20
TR1-565
M385
RP088-48(R)
RP3(MR)
R1028-KR(MR)
RP8(MR)
K10-5 (with risistant BB gene)
LSD0,01
LSD0,05

-4,742
-3,492
-4,325
-10,108

-2,425
4,958
7,858
6,858
3,008
3,192
8,879
6,541

Remark: By backcrossing and selecting the individuals ,the 9
promising lines have been selected, they have good characteristics of male line
such as high yield potential, good phenotype , high quantity of pollen. After
artificial screening for BPH only 5 lines have

moderate resistance to BPH ,

That is the RP8 line is resistant to brown plant hopper (score 3), the remaining
lines such as RP088-148, RP088-48, RP3, R1028-KR have medium resistance
grade (score 5). The new male lines with BPH resistance have a high general
combining ability value, respectively RP3 (7,858), R1028-KR (6,858), RPO8848 (4,958) and RP8 (3,008). these GCA are statistical significance at LSD0,05
level. The male lines do not carry the BPH resistance in general have low
general combining ability of the actual yield (negative value on GCA),
3.1.3. Results of breeding and selection of TGMS lines resistant to brown
planthopper
After the re-hybridization process and selection of sterile plants, in
2013 we selected 30 promising TGMS lines from some of the above hybrid
combinations. The lines of TGMS are compact plants, leaf colors from green to
dark green, long grains, only 4 lines are KR71-1, KR71-2, KR75-1 and KR31
had average seed size, the new TGMS lines are completely sterile in the 2013
crop conditions.

Test results of TGMS lines selected from KimS / E3 (KR67S) and
D116ST / E3 (KR95S) by RM6997 molecular markers have identified 5 plant


14
TGMS with BphZ gene from K67S and 2 plant TGMS carry BphZ gene from
KR95S (table 3.13)
The TGMS lines with BphZ will be selected on the genetic
background of KimS (AMS37S) and the genetic background of D116ST will
be a good material for selecting two-line hybrids with resistance to brown
planthopper Biotype 2 and Biotype3 in Vietnam.
Table 3.14. Results of identification of resistance genes for BPH Bph3,
TT
1
2
3
4
5
6
7
8
9
10

Bph6 and Bph9 in some new maternal lines were selected
Materials
Bph3 Bph6 Bph9 TT
Materials
Bph3 Bph6 Bph9
Rathuheenati

+
1 Rathuheenati
+
Swanalata
+
2
Swanalata
+
Pokali
+
3
Pokali
+
TN1
4
TN1
KR82-1
+
11
KR142-1
+
KR82-2
+
12
KR142-2
+
KR82-3
+
13
KR142-3

+
D59S-1
+
14
KR95-1
+
D59S-1-1
+
15
KR95-2
+
D59S -3
+
16
KR95-3
+
Source: Department of Biotechnology, FCRI; Note: +: Resistant homozygote
+/-: Heterozygous resistance;
-: infected;
0: not on the band
From the results summarized in Table 3.13 and 3.14, 5 lines of TGMS
containing BPH resistance genes and the following symbols are selected:
KR67S (KimS / E3), KR95S (D116ST / E3), KR142S (D52S / Pokkali),
KR82S (D59S / E2 // D59S) and D59S-1-1. Among them, two lines of TGMS
are KR95S (Bph3, BphZ), KR142S (Bph9) have many agronomic
characteristics of good maternal line such as: higher panicle exertion rate,
higher stigma exertion rate and longer panicle to use for further research.
Research results of some agro-biological characteristics of the TGMS
line KR95S and KR142S in the spring and summer crop seasion of 2017 in
Thanh Tri, Hanoi are presented in Table 3.17 (page 88). It show that: The two

TGMS lines KR95S and KR142S have growth duration in the short-day group,
the height of semi-dwarf plants. These characteristics are recorded in both of
the TGMS muntilication and F1 seed production Crops. The rate of stigma


15
exersion of the two lines is quite high at 75.5% (KR95S) and 73.6% (KR142S),
the KR95S line has a purple stigma and KR142S is white calour. Both of them
are typically sterile (WA), completely sterile temperatures are > 24.oC; The
ability of branching is quite good, the number of effective panicle reaches 7.9 9 panicle / hill, the number of flowers / panicle reaches 159 - 142 flowers /
panicle, grain shape of the two lines is average - long, the number of leaves /
main stem of KR95S is 13.5 and KR142S is 15, these two TGMS lines will be
good materials for breeding hybrid rice varieties with BPH resistance.
* Assessment of general combining ability (GCA) of new TGMS
Assessing the GCA of the TGMS lines to select the female lines with
high combining ability value, the selected female lines will have a higher
probability for the hybrid with superiority. Results of analysis for the GCA for
yield and yield components of 4 TGMS lines are presented in table 3.18,
Table 3.18. The general combining ability (GCA) of parents lines
For the yield components and productivity
Rate of
1000
Panicles/
Grains/
empty
grain
hill
panicle
Par.lines
grain

weight
Female (TGMS) lines
KR142S
0.29
-10.25
2.29
-0.06
D60S
-0.31
2.95
0.01
0.12
D67S
0.05
-3.05
-0.47
0.50
KR95S
-0.03
10.35
-1.83
-0.56
LSD0,05
0.05
2.50
0.10
0.15
LSD0,01
0.06
3.34

0.13
0.20
Male lines
SR3
-0.08
8.65
-0.74
0.06
SR14
0.12
7.4
-1.24
0.73
SR18
-0.18
6.65
-0.54
-0.12
AIQ6
-0.03
-3.35
1.17
-0.85
SR5
0.17
-19.35
1.34
0.18
LSD0,05
0.05

2.79
0.11
0.17
LSD0,01
0.07
3.74
0.14
0.22
Feature

Theoryield

Grain
yield

-1.56
-4.16
2.00
3.72
2.08
2.79

-0.625
-2.465
0.495
2.595
1.46
1.96

3.42

10.51
0.43
-5.51
-8.84
2.33
3.12

1.84
5.59
0.07
-2.79
-4.71
1.63
2.19


16
- Regarding the number of panicle/hill: The TGMS line -KR142S has
the highest general combining ability (0.29) and is significant at the level of
LSD0.01 (0.06),
- The number of grains / panicle: The KR95S having BPH resistance
gene has a very high GCA (10.35) and is significant at LSD0.01 (3.34), the
D67S line has a value of 2.9, the other two lines have negative values.
- The 1000 grain weight: KR59S and KR142S carrying BPH
resistance gene have negative value, while 2 lines D60S and D67S have values
of 0.12 and 0.5 are higher than expected meaning at the level of LSD0.01 (0.2).
- In terms of actual yield: There are 2 lines KR95S and D67S having
positive values for GCA. However, only KR95S have the highest positive
value of 2.595 being significant at the level of LSD0.01 (1,96).
3.2. The result of breeding and selection of TGMS lines resistant to

bacterial blight disease
3.2.1. Results of evaluation of materials for selection of parent lines
resistant to BLB
Evaluating 32 lines of genomic materials (NILs) carrying the blight
resistance gene (on the genetic background of IR24 rice variety) at the Field
Crops Research Institute showed that: The morphological characteristics of the
lines do not have much difference the lines, all have dark green leaves, Vcompact shape. Of the 32 lines with bacterial blight resistance genes, most of
them had head and beard in which IRBB13 line had the longest bearded head
(2.5cm). The grain color and granularity of the line samples containing
bacterial blight resistance genes are not significantly different, almost all lines
of grain color from light yellow to yellow and elongated granular form.
3.2.2. Results of transferring resistance to bacterial leaf blight into the
mother line TGMS
Electrophoresis the combinations with the M3 directive associated
with the Xa7 gene. Results of the V54 (AMS30S / IRBB7) electrophoresis
using the M3 indicator showed that V54-1, V54-2, V54-3, V54-4, V54-5 all
carried the Xa7 gene similar to the BB7 strain (Figure 3.7, page 100).


17
The study results showed that two Npb181 markers associated with
Xa4 and M3 genes linked to Xa7 gene for a clear polymorphic range were used
to identify the sterile line carrying the Bacterial leaf blight resistance genes. In
particular, using the Nbp181 directive identified 11 individuals of the V50
(AMS34S / BB60), V31 (AMS35S / BB60) and BL59 (AMS34S / BB60) lines
carrying the BLB resistance gene Xa4. Using the M3 directive, the V54 line
(AMS30S / IRBB7) was identified with 5 individuals carrying the Xa7 gene.
Comment: Throught screening uniformed TGMS lines, we selected
AMS34S similar TGMS lines containing Xa4 resistance gene, AMS35S similar
TGMS containing Xa4 gene and the TGMS line similar to AMS30S contains

the Xa7 gene. All these identified TGMS lines having bacterial leaf blight
resistance genes were improved from the original TGMS: AMS34S, AMS35S
and AMS30S, these original TGMS are being exploited as the mother line of
many hybrids in Vietnam. Therefore, new improved BLB resistant TGTMS
lines will be required firstly for crossing and selecting two line hybrid rice
varieties resistant to BLB in our country.
The lines of TGMS AMS35S-KBL and AMS30S-KBL have a
completely sterile temperature of ≥ 23.5 0C, stable sterility, high percentage of
stigmata exertion (70-75.8%), rice husk opening angle wide, the duration from
sowing to heading 10% in the summer season is 72 - 80 days, the degree of
BLB infection is 3-5 score in artificial conditions.
The lines of TGMS AMS35S-KBL and AMS30S-KBL have many
characteristics of good maternal flow (Table 3.22, page 103) selected by the
method of individual selection of sterile lines from BC4F2 generation.
* GCA assessment of yield of new TGMS lines with BLB resistance genes
Table 3.23. The combined ability value of some TGMS lines
Tester and line
GCA value
M415
-1,679
Tester (male)
TH29
1,679
LSD0,01
3,237
LSD0,05
2,337
Line (TGMS-female)
35S
0,474

18
36S
37S
D64S
D116ST
827S
35S-KBL
827S-KBL
LSD0,01
LSD0,05

-0,459
-0,681
-1,113
-2,979
1,967
0,521
2,271
5,027
3,629

The TGMS lines were assessed for their GCA and their specific
combining ability (SCA) for productivity by Line x Tester method and using
LINE * TESTER Ver 3.0 variance analysis software program by Nguyen Dinh
Hien (1996). Evaluation results of 8 lines TGMS are: 35S, 36S, 37S, D64S,
116 T, 827S, 35S-KBL, 827S-KBL with two test lines: M415 and TH29 (two
lines have been evaluated as two lines of females having a high degree of GCA
and a good line for hybrid rice 2 lines) showed that: The GCA of 35S-KBL and

827S-KBL has a higher positive value than other lines does not make sense at
the level of LSD0,05 but higher than the original TGMS lines
3.3. Hybrid rice breeding towards resistance to bacterial blight, brown
planthopper
3.3.1. Hybrid rice breeding towards resistance to brown planthopper
The results of evaluating the components of productivity and
productivity (Table 3.26, page 108) to estimate the potential of the variety, on
that basis,the growers can apply appropriate technical measures to maximizing
the advantages of each factor to gain better productivity for the crop.
Evaluating the number of effective panicle/hill of rice hybrids in the
Spring season. 2016 showed that most of the combinations have the average
number of panicle / hill from 5.0 - 7.0 panicle / hill, only the number 2
combination averaged 4.8 panicle /hill; The number of seed / panicle is quite
high and varies from 139 to 209 seeds / panicle, of which the highest number
of seeds / panicle is D60S / SR18 (209 seeds),


19
The combinations with female lines have the BPH resistant genes
have seeds / panicle from 159 - 204 seeds / panicle, control TH3-3 has 161
seeds / panicle.; The weight of 1000 grains of the combinations ranged from
23.4 to 28.9 grams, weight of 1000 grains of hybrids using female having BPH
resistance gene ranged from 24.8 to 28.7 grams, the control achieved 23.4
grams.
The theoretical yield of the combinations ranges from 9.52 to 13.6
tons / ha, the combinations with females carrying the BPH resistance genes
range from 9.52 to 12 , 14 tons / ha, control reached 9.04 tons / ha;
The actual yield of the combinations reached 7.67 - 9.76 tons / ha, the
combinations with females having BPH resistance gene had the actual yield of
7.73 - 9.32 tons / ha, much higher than the TH3-3 (7.06 tons / ha).

Results of pest monitoring (Table 3.27, page 109) show that: In the
Spring 2016 season, most of the combinations have the level of bacterial and
blight stripe infection from points 3-5.
Among 10 hybrids of female with the gene that was resistant to BPH
there were 2 hybrids (number 8, 9) infected with score 3, 5 hybrids (numbers 1,
12, 16, 17, 20) infected with score3-5 and 3 hybrids (numbers 4, 5, 13) infected
with score 5;
About BLB disease, there are 4 hybrids (numbers 1, 5, 8, 9) infected
with score 3, 4 hybrids (numbers 12, 16, 17, 20) infected with score 3-5 and 2
(number 4, 13) infections with score 5; Other pests such as blast disease do not
appear.
* Assessment of specific combining ability (SCA) of new TGMS lines with BPH
resistant gene:
Table 3.33. The specific combining ability of new TGMS lines
on actual yield
Females
KR142S

D60S

D67S

KR95S

Males
SR3

3.70

-2.16

1.08

-2.62

SR14

-2.85

-4.71

7.03

0.53


20
SR18

2.27

3.01

-1.55

-3.75

AIQ6

-3.78

2.26

-2.99

4.51

SR5

0.65

1.59

-3.57

1.33

LSD0,05

3.27

LSD0,01

4.38

The analytical results in the table 3.33, show that SCA of actual yield
of D67S / SR14 (7.03), followed by KR95S / AIQ6 (4,51) they are have
statistical significance at the level of LSD0.01 (4,38) and KR142S / SR3 (3.7)
has statistical significance at the level of LSD0.05 (3,27). 7 combinations have
positive values but no significant differences and 10 other combinations with

negative values
Hybrid with TGMS lines carrying BPH resistant genes and having
the highest SCA value in terms of the actual yield were hybrid KR142S / SR3
(3.7) gained (89.4 quintals / ha), anf the hybrid KR95S / AIQ6 (4,51) obtained
actual yield of 88.8 quintals /ha campared to 70.6 quintals / ha) of chech-TH33. It is expected that hybrds KR95S / AIQ6 and KR142S / SR3 will be most
promising hybrids
Conclusion : Through evaluation of hybrid combinations with the
participation of female lines carrying BPH resistance genes in the Spring
2016, There are 9 hybrids having actual yields from 85.1 - 93.2 quintals / ha
being higher than control TH3-3 (70.6 quintals / ha) 20% or more. However,
only 6 hybrids have TGMS lines carrying BPH resistant genes combining with
high yields were selected for further yield trials in next season .The selected
hybrids are KR142S / SR3 (89.4 quintals / ha) , KR142S / SR14 (86.6 quintals
/ ha), KR142S / SR18 (86.2 quintals / ha), KR95S / SR14 (93.2 quintals / ha),
KR95S / AIQ6 (88.8 quintals / ha) and KR95S / SR3 (86.3 quintals / ha).
3.3.2. Hybrid rice breeding towards resistance to bacterial blight
Assessing the degree of BLB damage of hybrid combinations in the
crop condition by artificial infection in the field (Table 3.35, page 119) shows
that: Most of the hybrids usingTGMS lines are resistant to bacterial leaf blight


21
at score 5-7, while hybrids having female with BLB resistance genes have a
lower degree of infection. TGMS line AMS34S-KBLis used as female of 2
hybrids (numbers 7, 9) have high resistance score 1-3,one hybrids(number 6,
8) have resistance score 3 and one hybrid (numbers 10) have medium
resistance -score 3-5. The female line AMS35S-KBL has 1 hybrid with BLB
high resistance- score 1-3 (number 29), 1 hybrid having score 3 (number 27)
and 3 hybrids having medium resistance- score 3-5 (numbers 26, 28, 30); The
mother line AMS30S-KBL has 3 hybrids resistance - score 3 (number 16, 19,

20) and 2 hybrids have medium resistance- score 3-5 (number 17, 18); the
check have moderate resistance -score 5.
Results of evaluation of yield components and yield of hybrids in
(Table 3.36) show that: number of panicle / m2 of combinations ranges from
224 - 315 panicle / m2 (Number 24), in which there are 3 combinations
(numbers 10, 12, 24) with the number of panicle / m2 being higher than 270
panicle/ m2, 6 combinations (numbers 8, 14, 17, 18, 22, 28) reaching 250 - 266
panicle / m2.
The number of grains / panicle of hybrids using AMS34S-KBL as
female ranged from 121 to 165 seeds / panicle being higher than those of
mother AMS34S (114-125 seeds / panicle); The hybrids using the AMS30SKBL have the number of filled grains / panicle varying from 114 to 130 grains
lower than the hybrids of AMS30S (ranging from 119 to 161 seeds); The
hybrid of the AMS35S-KBL have a number of filled grain / panicle ranging
from 118 to 150 seeds, higher than the hybrid combinations of AMS35S (98 136 seeds). In the 2016 crop, the sterile seed rate of the hybrid are quite high,
ranging from 10.2 to 23.8% The weight of 1000 grains of the hybrids fluctuates
from 20.8 - 28.1 grams (number 30), the control TH3-3 is 23,7g.
- The theoritical productivity of the combinations ranges from 64.6
(number 17) - 10,08 tons / ha (No. 28 - AMS35S-KBL / R100).
- The highest actual yield is 72.1 quintals / ha of AMS35S-KBL /
R100 (HYT124-KBL) compared the yield of 67.8 quintals /ha of AMS35S /
R100 named HYT 124; The combination AMS30S / R116 is actually named


22
HYT 116 gained actual yield of 67.3 quintals / ha compared to combination
AMS30S-KBL / R116 (HYT116-KBL) reaching 69.4 quintals / ha,
Table 3.42. Specific combining ability (SCA) of new TGMS lines on
Females
Males

AMS34S

productivity (actuall Yield)
AMS34SAMS30SAMS30S
KBL
KBL

AMS35S

AMS35SKBL

SR14

2.43

1.37

-4.83

3.97

-0.81

-2.11

RP3

2.48

-0.78

1.12

-4.98

5.74

-3.56

R100

-3.36

-5.42

-5.62

0.48

4.50

9.40

SR18

0.93

5.97

4.37

-6.53

-2.51

-2.21

R116

-2.47

-1.13

4.97

7.07

-6.91

-1.51

LSD0,05

2.37

LSD0,01

3.16
Selecting 3 combinations with high SCA value of high actuall yield

are: AMS35S-KBL / R100 (HYT124-KBL) giving the highest SCA value of
9.4, combination of AMS30S-KBL / R116 (HYT116-KBL) is 7.07 and
AMS34S-KBL / SR18 achieved SCA value of 5.97.
Final results of our BLB breeding programme:
Through breeding and evaluation of hybrid combinations in the
direction of resistance to BLB in the season of 2016, we selected combinations
with higher yield than check variety > 10% (yield> 6.5 ton / ha) are AMS35SKBL / R100, AMS30S-KBL / R116 and AMS34S-KBL / SR18 in which the
most productive combination is AMS35S-KBL / R100 (called HYT 124-KBL)
reaching 7.21 tons / ha, bacterial leaf blight resistance score 3-5; The second
combination is AMS30S-KBL / R116 (called HYT 116 - KBL) with actual
yield of 6.94 tons / ha, BLBresistance score 3 compared to 5.95 tons / ha and
resistance to BLB resistant score 5. of check TH3-3
The evaluation also showed that the two improved hybrids
HYT124-KBL and HYT116 - KBL have similar

viz,

characteristics of the

original hybrids, but the ability to resist bacterial leaf blight are significantly
improved. (table 3.43, 3.44- page 128, 130)


23
CONCLUSIONS AND RECOMMENDATIONS
1. Conclusion:
1- Created 2 new TGMS lines having BPH resistant gene: KR95S
have BPH resistant genes Bph3, BphZ and KR142S having gene Bph9. These
TGMS have short growth duration in the summer season, pollen sterile
stability, critical temperature to induce complete pollen sterility at >24 oC, ratio

of stigma exertion >70 %. KR95S have GCA for traits of grain number/panicle,
theory yield and actual yield. These TGMS lines are good female parents for
breeding 2 line hybrid rice to tolerate to BLB and BPH in Vietnam.
2- Created 5 new male parental lines tolerant to BPH (from medium
resistance to high resistance), having good characteristics such as high yielding
capacity, good phenotype, high pollen production. These lines are RP8, RP08848 resistant to BPH (score 3); RP3 having gene Bph3 resistant to BPH
(score5); R1028KR tolerant to BPH (score5). Through screening male
papulation by the molecular markers, the individual plants of R116 having
gene bph6 was sellected for use. Four among five of the new BPH resistant
lines have high general combining ability (GCA) for grain yield following in
this order: RP3, R1028-KR, RPO88-48 and RP8.
3- Created 2 new TGMS lines having BLB resistant genes: AMS35SKBL (having Xa4) and AMS30S-KBL (having Xa7), these two lines have
critical temperature to induce complete pollen sterility at >23.5oC, pollen
sterile stability, ratio of stigma exertion (70 %-75.8%), level of BLB tolerance
at artificial conditions (score3-5). AMS35S-KBL have positive value of GCA
fot actual yield.
4- Breeding 3 hybrids used female-TGMS having BLB resistant genes
and resistant to BLB (score 3-5), having actuall yield over check to be more
than 10 percent. They are AMS35S-KBL/R100, AMS30S-KBL/R116 and
AMS34S-KBL/SR18, among them AMS35S-KBL/R100 (named HYT124KBL) gained highest yield (7.1 tons/ha). Second hybrid is AMS30SKBL/R116 (named HYT116 – KBL)

have actuall yield of 6.94 tons/ha,

resistant to BLB (score3) compared to yield of check (5.95 ton/ha) having
resistant to BLB (score5)