Hybrid Rice Breeding & Seed Production

FANGMING XIE International Rice Research Institute DAPO BOX 7777 Metro Manila, Philippines f.xie@cgiar.org

What is Hybrid Rice?

The first generation offspring of a rice cross between two genetically diverse parents

How Hybrid Rice?

Normal Rice Spikelet

(self pollinated crop)

Sterile Rice Spikelet

(Male Sterility)

Hybrid Seed Production

(Male Sterile x Normal Rice)

Why Hybrid Rice?

Heterosis (Hybrid vigor) Application to Increase:
Productivity (yield/unit/time, 15-20% of yield advantage), and Economic returns

Heterosis
A universal phenomenon that F1 generation shows superiority to both parents in agronomic traits or yield It presents in all biological systems and has been exploited commercially in many agricultural crops.

How to Measure heterosis?

Mid-Parent (MP) heterosis (F1 performs better than mean of two parents):
F1-MP X100 MP

Better Parent (BP) heterosis (F1 performs better than better parent):

F1-BP X100 BP

Standard heterosis* (F1 performs better than the check variety):

F1-CK X100 CK

* Standard heterosis is the most useful term in commercial crop production

Male Sterility Systems in Rice

Male sterility: a condition in which the pollen grain is unviable or cannot germinate and fertilize normally to set seeds. Male Sterility Systems (genetic and non-genetic):
Cytoplasmic genetic male sterility (CMS) Male sterility is controlled by the interaction of a genetic factor (S) present in the cytoplasm and nuclear gene (s). Environment-sensitive genic male sterility (EGMS) Male sterility system is controlled by nuclear gene expression, which is influenced by environmental factors such as temperature (TGMS), daylength (PGMS), or both (TPGMS). Chemically induced male sterility Male sterility is induced by some chemicals (gametocides)

Brief history of hybrid rice

1926 - Heterosis in rice reported 1964 - China started hybrid rice research 1970 - China discovered a commercially usable genetic tool for hybrid rice (male sterility in a wild rice = Wide Abortive ) 1973 - PTGMS rice was found in China 1974 - First commercial three-line rice hybrid released in China 1976 - Large scale hybrid rice commercialization began in China 1979 - IRRI revived research on hybrid rice 1981 - PTGMS rice genetics and application was confirmed 1982 - Yield superiority of rice hybrids in the tropics confirmed (IRRI) 1990s - India and Vietnam started hybrid rice programs with IRRI 1991 - More than 50% of Chinas riceland planted to hybrids 1994 - First commercial two-line rice hybrid released in China 1994 - 1998 - Commercial rice hybrids released in India, Philippines Vietnam

Rice and Hybrid Rice Production in China

40 35 30 64 56 48 40 32 24 16 8 0 1976 1978 1980 1982 1984 1986 1988 1990 1992 1994 1996 1998 2000 2002

25 20 15 10 5 0

Year
Total R ice Area Hybrid Rice Area % of Hybrid rice Area

% of hybrid rice area

Area (million ha)

Rice Grain Yield in China

Rice Grain Yield in China 7500 7000 6500
Grain yield (kg/ha)

6000 5500 5000 4500 4000 3500 3000 1976 1978 1980 1982 1984 1986 1990 1992 1996 1998 2000 2002 1988 1994

Year Hybrid rice Inbred rice

Hybrid Rice Area in other Asia Countries

Country
1997 Bangladesh India Indonesia Myanmar Philippines Vietnam Total 188 288 2 13 480 710 100

Hybrid Rice Area (1,000 ha)

2001 15 200 2004 50 560 10 42 189 650 1,445 2005 90 NA NA NA 367 NA 300(DS) 2006 150 (Exp)

Hybrid Rice Production in Vietnam

700 600 7.4 6.8 6.2 5.6 5.0 4.4 3.8 3.2 1997 1998 1999 2000 2001 2002 1992 1993 1994 1995 1996 2003

Area (1,000 ha)

500 400 300 200 100 0

Year
Are a (1000 ha ) Hybrid Rice Yie ld (t/ha ) Na tiona l a ve ra ge rice yirld (t/ha )

Yield (t/ha)

Two Commercial Systems for Hybrid Rice

Requirements for 3 Lines in CMS System

A-line Stable Sterility Well developed floral traits for outcrossing Easily, wide-spectum, & strongly to be restored B-line Well developed floral traits with large pollen load Good combining ability R-line Strong restore ability Good combining ability Taller than A-line Large pollen load, normal flowering traits and timing

TGMS and two-line hybrid

Temperature

high

Reproductive Upper Limit

Sterile F1 Seed Production

Based on the discovery of P(T)GMS mutant Male sterility controlled by 1 or 2 pairs of recessive gene(s)
low

Critical Sterility Point

Partial Sterility

Critical Fertility Point

Fertile S-line Multiplication

Reproductive Lower Limit

Model of Sterility / Fertility Expression for TGMS Rice

Flow chart of 3-Line H ybrid R ice E valuation and S eed P roduction

Elite C M S line SO UR C E NU RS ERY T o evaluate parents and m ake testcross Elite lines from different sources B & R line B reeding Program P line Breeding Progam CM S B AC KCR O SS N UR SERY BC2- BC4, C MS Evaluation TESTCRO S S NUR SERY T o identify B, R & P lines Backcross C M S pairs (BC 1) Prem arily heterosis evaluation, 2 rows w/ parent AxB Paircross B reeder Seeds RETE STCR O SS N U RSE RY (O YT) Re-evaluate F1 hybrids S tage 1, 1 rep, 3 rows AxB Increase Core Seeds AxB Seed Production Foundation Seeds AxB Seed Production Certified Seeds A & B Line Release Prelim inary Yield Trial (PY T) Stage 2, 1 rep, plot Hybrid S eed Production for AYT & N Y T Advanced Y ield Trial (AYT) Stage 3, 3 reps, plot N ational Yield Trial Stage 4, 3-4 reps, m uti-location, 2-years O n-Farm Trial (S trip Trial) H ybrid P ilot Seed Production Isolation B lock Hybrid and R line R elease Isolation B lock Hybrid Seed Production for PY T Isloated Net or bags H ybrid Seed Production for O YT Isolation B ags or hand-crossing R & P Line

F lo w c ha rt o f 2 -L in e H yb rid R ice E va lu a tio n an d S eed P ro d u ctio n

SO URCE NURSERY TG M S L in e B reed in g T o e valua te paren ts and m ak e testcross E lite lines from d ifferent source s B & R lin e B reed ing P rog ram P ollinato r lin e B ree ding P rog am B re eder S ee ds C o re S e eds F ound ation S ee d C ertified S eed s TG M S Lin e R elease TE S TC R O S S N U R S E R Y T o identify T G M S & P lines P re m arily heterosis evaluation , 2 row s w / parent R E TE S TC R O S S N U R S E R Y (O Y T) R e-evaluate F1 hybrids S tag e 1, 1 rep, 3 row s P relim ina ry Y ield Trial (P Y T) S tag e 2 , 1 rep, plot Advan ced Y ield Trial (AY T) S ta g e 3, 3 reps, plo t H ybrid P ilot S eed P ro d uc tion N atio nal Y ield Tria l S tag e 4, 3-4 re ps, m uti-location, 2-years H yb rid an d R lin e R eleas e O n -Farm Tria l (S trip Trial) Isolation B lock H yb rid S eed P ro duction for AY T & N Y T Isolation B lock H ybrid S eed P rod uction fo r P Y T Isloated N et or bag s H yb rid S eed P rodu ctio n for O Y T Isolatio n B ag s o r han d-cro ssing

Advantage & Disadvantage of 3-line hybrid rice system

Advantages Stable male sterility Disadvantages Limit germplasm source (CMS, Restorer) Dominant CMS cytoplasm in large area (WA) One more step for parental seed production Time consuming of CMS breeding

Advantage & Disadvantage of 2-line hybrid rice system

Advantages Simplified procedure of hybrid seed production Multiple and diverse germplasm available as parents Any line could be bred as female 97% (2-line) vs 5% (3-line) of germplasm as male Increased chance of developing desirable & heterotic hybrids Multiple cytoplasm courses as female parents Disadvantages Environmental effect on sterility could cause seed purity problem

Two-line hybrid production in China

Tw o Line Hy brid Rice In China
7800 7600 7400 1600 1400 1200

7000 6800 6600 6400 6200 1991 1993 1995 1997 2002 1992 1994 1996 1998 1999 2000 2001 2003

800 600 400 200 0

Year
2-line Hybrid Yield A ll Hybrid Yield Area

Area (1000 ha)

Yield (kg/ha)

7200

1000

Hybrid Rice Seed Standard

H y b r id R ic e S e e d S ta n d a r d (G B 4 4 0 4 .1 - 1 9 9 6 , C h in a )

Seed S t e r ile L in e M a ia n t a ie r R e s to re H y b r id

C la s s C o re

P u r it y C le a n lin e sG e r m in a t io n M o is tu r e s ( > %) ( > %) ( > %) ( < %) 9 9 .9 8 5 .0

1 3 .0 ( in d ic a ) 1 4 .5 ( ja p o n ic a )

F o u n d a tio n9 9 .0 1st 2nd 9 8 .0 9 6 .0

9 8 .0

8 0 .0

1 3 .0

Mission of IRRI Hybrid Rice Program

Developing germplasm, parents and hybrids as internationally public goods Research new technology for breeding and seed production Collaboration with NARS and private sectors in hybrid rice research and production Promotion of exchange of information, technology, scientist and germplasm

Strategy of IRRI Hybrid Rice Program

Focusing on conventional tools and integrate them with proven non-conventional methods to develop the technology Developing parental lines, especially female parents with high outcrossing and high quality, to promote hybrid rice spreading Facilitating development of close partnership between public and private sectors in national programs Intensifying agronomic research to get maximized manifestation of heterosis in hybrids

Release of IRRI Hybrids in Different Countries (1994-2005)

IRRI Hybrid IR64610H IR64611H IR64616H IR65489H IR68284H IR69690H IR69690H IR69690H IR69690H IR75207H IR75217H IR78386H Released as MGR-1 KRH-1 Magat DRRH-1 Mestizo 1 Sahyadri HYT-57 BRRI Dhan Hybrid 1 Rokan Mestizo 2 Mestizo 3 Mestizo 7 Country India India Philippines India Philippines India Vietnam Bangladesh Indonesia Philippines Philippines Philippines Year released 1994 1994 1994 1996 1997 1998 1999 2001 2002 2002 2002 2005

Release of Hybrids by using IRRI Germplasm in Different Countries (1994-2004)

Hybrids released by NARS using IRRIbred CMS lines
Hybrid name APHR-1 APHR-2 CNRH-3 KRH-2 Pant Sankar Dhan-1 ADTRH-1 CORH-2 Narendra Sankar Dhan-2 Rokan Maro Hipa 3 Hipa 4 Country India India India India India India India India Indonesia Indonesia Indonesia Indonesia Year released 1994 1994 1995 1996 1997 1998 1998 1998 2002 2002 2004 2004 HR 120 (6444) India 2001

Hybrids derived from IRRI-bred parental lines and commercialized by private sector
Hybrid name Biganti Intani 1 Intani 2 PHB-71 Proagro 6201 Country Philippines Indonesia Indonesia India India Year released 2004 2001 2001 1997 2000

Germplasm Shared

Super high-yielding hybrid rice breeding in China

P ro g r e s s o f " S u p e r H ig h -Y ie ld in g " H y b rid R ic e P r o g ra m in C h in a
G o a l (s in g le -s e a s o n ) S ta g e S ta rt Phase I P h a s e II P h a s e III Y ie ld (t/h a ) Y e a rs 1997 1996 - 2000 2000 - 2005 2005 - 2010 F in is h e d F in is h e d S ta r te d L a r g e a r e a e x te n s io n S ta r te d P r o g r e s s (2 0 0 6 ) B r e e d in g C o m m e r c ia liz a tio n

8 .2 5 1 0 .5 1 2 .0 1 3 .5

Morphological Model of Super High-yielding Hybrid Rice

Plant height = 100 cm, with culm length = 70 cm Uppermost three leaves: Flag leaf, long, 50 cm, higher than the panicle top 20 cm. The 2nd leaf from the top: 10% longer than the flag leaf, and over the top of the panicle. The 3rd leaf = the middle position of the panicle Erect: the leaf angles of the flag, 2nd and 3rd leaves are around 5, 10, 20 degrees, till mature Narrow, V-shape and thick: narrow with 2 cm when flattened. Plant type: moderate compact with moderate tillering capacity; drooping panicles after filled, above ground ~ 60 cm, erect-leaved canopy without appearance of the panicles Panicle weight and number: grain weight per panicle = 5 g, 2.7 million panicles per hectare. Leaf area index (LAI) and ratio of leaf area to grains: the LAI is ~ 6.5 based on the uppermost three leaves, the ratio of leaf area to grain weight is 100 : 2.2-2.3, meaning that to produce 2.2-2.3 grams of rice, 100 cm2 of the upper three functional leaves are needed. Harvest index > 0.55

Hybrid Heterosis in Rice

Indica x japonica Indica x javanica japonica x javanica indica x indica japonica x japonica

Inter-subspecific hybrid rice breeding

Difficult in breeding inter-subspecific hybrid rice Low seed set Tall plant height Poor grain-filling Late maturity Grain quality market Solution for breeding of inter-subspecific hybrid rice wide compatibility (WC) genes allelic dwarf gene indica/javanica hybrids in indica rice growing region japonica/javanica hybrids in japonica rice growing

Future Opportunity: Enhance yield heterosis

Exploiting subspecies heterosis Applying biotechnology for parent selection (heterotic groups and/or heterotic gene blocks Enhanced yield heterosis in indica / NPT hybrids compared to indica / indica hybrids (retestcross, IRRI, 2004WS)
Hybrid # of heterotic hybrids Total # of hybrids % of heterotic hybrids Yield advantage (%) over best inbred check Range Mean

indica / indica

34 20

85 40

40 50

1-80 6-131

29 42

Indica / NPT

Future Opportunity:
Increase yield of hybrid seed production
Performance of hybrid seed production in tropical countries (2003-2004)

Yield (kg/ha)
Country
India Vietnam Philippines Bangladesh Indonesia China

Mean
1,600 2,000 810 (04DS) 800 500 2,750

Range
1,000 4,500 1,500 3,500 600 2,000 600 2,000 300 1,600 1,500 6,000

Developing high outcrossing parents Improving seed production technology Training seed growers Selecting adequate location / season

Hybrid Rice Seed Production in China

Hybrid Rice Seed Production in China
3500 3000 350 300

2000 1500 1000 500 0 1978 1980 1982 1984 1988 1990 1992 1994 1998 2000 1976 1986 1996 2002

200 150 100 50 0

Year
Yield Area Linear (Yield) Linear (Area)

Area (1000 ha)

Yield (kg/ha)

2500

250

Hybrid Rice Seed Production

In Asia In United Sates

Future Opportunity:
Improve hybrid rice grain quality
Measurements of Rice Grain Quality: Milling yield Total milling yield Whole milling yield Chalk Amylose content Gel Temperature (ASV) Length, width, L/W Protein Aroma

Future Opportunity:
Improve hybrid rice grain quality
Rice grain quality of inbreds and hybrids*

Trait
Total Milling (%) Whole Milling (%) Chalk (%) Amylose (% GT Length L/W

Inbred
69.1 48.7 13.5 19.8 4.3 6.9 3.2

Hybrid
68.2 45.4 20.6 20.6 5.5 7.1 3.3

Data from National Cooperative Testing (NCT), Philippines, 2004-2005

Difference of Whole Milling Yield and Chalk between Inbreds and Hybrids
Whole Milling Yield and Chalk in Hybrids and Inbreds (NCT, 2004-2005, Philippines)
60

Average Whole Milling (%) Hybrid = 45.4 Inbred = 48.7 Average Chalk (%) Hybrid = 20.6 Inbred = 13.5

50

40

Chalk (%)

30

20

10

0 20 25 30 35 40 45 50 55 60 65

Whole Milling Yield (%) Hybrid Inbred

Difference of Whole Milling Yield and Chalk between Inbreds and Hybrids
D is tr ib u ti o n o f W h o l e M i ll i n g Y ie l d
D i s tr i b u ti o n o f C h a l k

40 35 30 25 % of Entries
% of Entries

45 40 35 30 25 20 15 10 5

20 15 10 5 0 <30 30-35

H y b r id ( n = 1 1 3 ) 0 35 -4 0 40 -4 5 4 5 -5 0 In b r e d ( n = 1 6 ) 50-55 >55

<5

H y b r id ( n = 8 0 ) 5 -1 0 10 -1 5 15-20 In b r e d ( n = 1 4 ) 20-25 >25

M illin g Y ie ld R a n g e

C h a lk R a n g e

Data source: 2004 and 2005 NCT, Philippines

Future Opportunity:
Develop hybrids for unfavorable environments
Hybrids Have Substantially Improved Yield under Severe Lowland Stress (ca. 1 t/ha). (IRRI, G. Atlin, 2005)
IRRI DS 2004-5
8000

IRRI DS 2005
5000

18.1%

7000 6000 Grain Yield (kg/ha)

Yield advantage 10.5%

1.9%

4500 4000 Grain Yield (kg/ha) 3500 3000 2500 2000 1500 1000 Non-stress Environm e nt Hybrid (n=3) Inbred (n=7) Stress

5000 4000

49.8%

78.6%
100% 100%
Full

3000 2000

83% 89%
A lternate Wetting & drying Ir r igation M e thod Hybrid (n=2) Inbred (n=6)

43% 29%
Drought

100%

67% 46%

1000

100%

Future Opportunity:
Improve agronomic management and deployment strategy
Unhealthy Healthy canopy

ShanYou 63 grown under different nitrogen management (S. Peng, IRRI)