ACHIEVEMENTS AND DEVELOPMENT OF HYBRID RICE IN CHINA Dr. Ma Guohui Deputy Director General China National Hybrid Rice R & D Center July, 2, Bangkok

Overview: Rice Production Area: 30 Mil ha yearly (33.88Mil ha in 1980) Average Yield: 6.16~6.78 ton (2001~2012). Output: returned the high level up to 200 million tons in 2012

Overview: Hybrid Rice Contribution Basic Chinese situation ● population: about 1.3 billion ● population rate in the world: 22% ● arable land rate in the world: 7% China became self-sufficient in basic food for the first time in modern history Tremendous accomplishment in last century, the innovation of HYBRID RICE, made by Prof. Yuan Longping

Overview: Hybrid Rice Contribution China is the first country to commercialize hybrid rice Yearly area: about 17.6 mil. ha, 54% of total rice area Yield: 7.5t/ha of HR with 20% yield advantage over inbred Yearly production of HR: about 120 Mil. tons, about 57.5% of the total rice production Up to year 2013, the accumulated planting area of HR is 380 mil. ha and the increased paddy by HR is about 600 mil. tons Yearly area Mil ha Yearly production: Mil tons Total area and production of HR Inbred Hybrid Inbred 42.5% Inbred 46% Hybrid 57.5% 54% total rice area total rice production

Overview: Hybrid Rice Contribution Seed enterprises (in 2012) 520 Seed enterprises which have the capacity to the breeding, seed production and marketing for HR production The commercial seeds of HR 283 million kg/y Employment opportunities Direct: 0.1 million/y Indirect: 1.0 million/y

1. History of hybrid rice in china 1) Three-line HR Development In 1964, Prof. L.P. Yuan initiated HR research in China. In November 1970, a pollen abortive wild rice (WA) discovered. In 1972, Prof. Yuan developed the first CMS line: Erjiunan 1A . In 1974, The first rice hybrid Nanyou No.2 developed by Prof. Yuan. In 1980, Shanyou63 developed by Prof. Xie, the largest one of growth area in 1980's Completion of the three-line hybrid rice system within 10 years

2) Two-line HR Development In 1973, the first natural male sterile rice plant discovered by Mr. S.M. Shi. A new way to utilize the heterosis in HR by two-line system with PTGMS. In 1986, the strategic breeding approach by Prof. Yuan: on the methodology from three-line, two-line to one-line, and on the level of heterosis from inter-varieties, inter- subspecific to distance hybrids. Bw 1987 to 1995, as the principal scientist, Prof. Yuan developed a comprehensive array of theories of “two-line system hybrid rice”

(reproducing parent seed） (producing hybrid seed) The fertility conversion theory and mode of PTSG lines established by Prof. Yuan Fertility (reproducing parent seed） Critical sterile T Biological Max T Biological Min T Sterility (producing hybrid seed) Partial sterility Critical fertile T 在两系法杂交水稻育种中，我们首次在籼稻中发现光温敏不育材料安农S，同时探明了光温敏核不育系的温光作用模式，提出了选育实用光温敏不育系以及核心种子繁殖的理论与技术，育成了第一个实用光温敏核不育系培矮64S。 安农S和培矮64S及其衍生系已成为两系法杂交稻的主流亲本，是两系法杂交稻最具实用价值的原创骨干亲本。 而培矮64S更是全国应用最广、推广面积最大的两用核不育系，其系列组合累计种植面积1亿多亩，为我国两系法杂交稻应用起了十分关键的作用。 First viable TGMS: Peiai64s The fertility conversion mode of PTSG sterile lines 8

The success of two-line hybrid rice research was declared in 1995. Area of two-line HR: 3 million ha in recent years High yield potential with good quality, 5~10% higher than three-line HR

2. Progress in HR development 1) The commercial HR production and varieties release The stage of rice development: Before 1964 – T.V. stage: about 2 t/ha 1965 ~ 1976 - SSD stage: ≈about 3.5 t/ha 1976 to now - HR stage: ≈ 5~6.3 t/ha The average yield of HR in 1990s:7.5 t/ha

Varieties release of commercial HR Types of rice variety 2012 2011 2010 2009 2008 Number % Indica inbred 30 7.6 33 8.6 32 6.4 36 7.5 25 5.6 Indica hybrid 247 62.8 257 67.2 313 63.3 303 63.7 291 65.2 Jalonic inbred 102 25.9 77 20.1 122 24.7 107 22.5 113 25.3 Jponic hybrid 14 3.5 15 3.9 27 5.4 29 6.1 17 3.8 Total 393 100 382 494 446 ● 5883 of rice varieties released during 2008 ~ 2013 ● About 400 varieties released yearly ● HR varieties released: 66.3~71.1%

Yield standard of the Super Rice in China 2) Development of super HR Super rice program launched by MOA of Chinese in 1996 Yield standard of the Super Rice in China Phase Hybrid Rice Yield increase(%) First cropping Second cropping Single cropping 1996 level 7.50 8.25 Phase Ⅰ 1996-2000 9.75 10.50 More than 20% Phase Ⅱ 2001-2005 11.25 12.00 More than 40% Phase Ⅲ 2010-2015 / 13.50 More than 60% Phase Ⅳ 2015-2020 15.00 More than 100% * Tons/ha at 2 sites with 6.7 hectares in each site in 2 consecutive years.

The real ideal model of plant morphology of hybrid (Pei’ai 64S/E32) Super HR Program proposed by Prof. Yuan Morphological Improvement Upper 3 leaves: long, erect, narrow, V-shape and thick Height of canopy: 1.2 m above The real ideal model of plant morphology of hybrid (Pei’ai 64S/E32)

Raising Heterosis Level Seed-setting rate(%) Raising Heterosis Level The heterosis utilization theory of indica-japonica subspecies proposed by Prof. Yuan I/JP I/JV JP/JV I/I JP/JP Spikelets /p Dry matter weight per plant (g) Indica (I) / japonica (JP) / Javanica (JV) Heterosis in Different kind of Rice Hybrids

Super hybrid rice breeding theory and strategy To be the higher plant based on drawf gene To develop the nearer parent from far genetic distance relationship Use of dominance combines with super-dominance Panicle to be between moderate and large Higher ratio of grain to leaf Grain plumpness based on well grain filling parents Achieve good grain quality based on Javanica To be well ecological adaptation. 为了培育优势更强的杂交稻组合，我们研究并提出了籼粳亚种间杂种优势利用的理论与技术，基本策略是32个字：矮中求高、远中求近、显超兼顾、穗中求大、高粒叶比、以饱攻饱、爪中求质、生态适应。 运用这种策略创制了培矮64、零轮等广亲和系；利用籼粳亚种间杂交育成长江流域推广面积最大的组合金优207。 15

Utilize the Biotechnology I: Utilization of favorable genes from wild rice II. Using Genomic DNA From Barnyard Grass to Create New Source of Rice Ⅲ: Using C4 gene from maize to transfer into super hybrid rice

A draft sequence of super hybrid rice parents completed firstly in the world It promotes the understanding of rice heterosis at the genomics level and molecular breeding applications in HR

生命科学权威刊物Cell在其"Leading Edge"专栏中专文介绍并给予高度评价 Major breakthroughs were made in molecular mechanism research of rice heterosis in 2009 Using gene chip technology to carry out the super hybrid rice LYP9 and its parent transcriptome studies, identify potential associated with super hybrid rice heterosis functional genes. “Cell” commented: a new starting line to elucidate the molecular mechanisms of rice heterosis 2、水稻杂种优势的分子机理研究取得重大突破 利用基因芯片技术开展了超级杂交稻LYP9及其亲本的转录组学研究，鉴别出与超级杂交水稻杂种优势相关的潜在功能基因。 相关研究论文发表在美国PNAS杂志上，这项研究被评为2009年度中国基础研究十大新闻。 生命科学权威刊物Cell在其“前沿”专栏中专文介绍了这项研究，并给予高度评价，认为该项研究是阐明水稻杂种优势分子机理的一个起点。 Using gene chip technology to carry out the super hybrid rice LYP9 and its parent transcriptome studies, identify potential associated with super hybrid rice heterosis functional genes. Related research papers published in the American journal of PNAS, this study was chosen as the basic research of 2009 year China ten big news. Life science authoritative journal Cell in the "frontier" column article introduces the research, and give high praise, think the study is to elucidate the molecular mechanisms of rice heterosis a starting point. 生命科学权威刊物Cell在其"Leading Edge"专栏中专文介绍并给予高度评价 18

Molecular marker assisted selection (MAS) Identified and mapping genes of QTL (yld1.1 and yld2.1) from wild rice (O.r ufipogon) and published in “Nature”; Developed an elite restorer lines and hybrid rice combinations with yield increase of 20%, such as Y liang-you 7 3、首次发现野生稻增产QTL并进行分子标记辅助育种 在野生稻中发现了增产QTL（yld1.1和yld2.1），其增产效应分别达到18.26%和17.07%。研究论文发表在《Nature》杂志上。 通过分子标记辅助选择，培育出强优恢复系及 Y两优7号等超级杂交稻组合。 “野生稻高产基因分子育种技术体系及其应用研究”获得湖南省科学技术发明一等奖。 Nature 384:223-224 19

Favorable genes from barnyard grass transferred and developed specific parents for super hybrid rice Zi100B Oryza minuta V20B 4、转移野生稻、稗草的远缘有利基因，培育出超级杂交稻特异亲本品系及组合 为阐明远缘物种间遗传信息转移的规律，开展了分子机理方面的研究，包括蛋白组学、 DNA序列分析等等。获得了供体物种大片段DNA转移的分子证据，发现了外源DNA导入引起受体发生突变的“热点”现象。部分研究成果已发表在植物学报、蛋白组学研究等杂志。 To clarify the law of the transfer of genetic information between distant species, carried out the mechanism of molecular research, including proteomics, DNA sequence analysis, and so on. The molecular evidence of large fragments of DNA transfer donor species, found the exogenous DNA into receptor mutations caused by the phenomenon of "hot spots". Some research results have been published on JIPB and J. Proteome Res. RB207 R207 barnyard grass J. Proteome Res., 2007, 6:1354–1363 20

C4 rice: get transgenic lines to be used to HR breeding PEPC PPDK NADP-ME MDH Transform hybrid rice parents The production of transgenic lines Line Transfrom genes Compared with CK production increase(%) Fengyou299 CK - FengA/09T005 PEPC+PPDK 8.60% FengA/09T009 MDH+ME 1.86% FengA/09T015 PEPC+PPDKM+DH+ME 3.78% C4植物比C3植物的光能利用率高30%，我们正在将C4植物玉米的一些相关基因转移到C3植物水稻，通过创制C4型水稻来提高产量潜力。玉米C4基因已被克隆，成功导入超级杂交稻亲本中，获得光合效率大幅度提高的中间材料。 The light energy utilization rate of C4 plants is 30% higher than C3 plants, we transformed the relevant genes from maize transferred to C3 rice, through the creation of a type C4 rice to increase yield potential. C4 genes have been cloned successfully and imported super hybrid rice parents, photosynthetic efficiency increased significantly in the intermediate materials.

Great Progress of Super HR 15t/ha (14.82) 12t/ha 10.5t/ha Phase 4 Phase 3 Phase 2 Phase1 In 2020 In 2015 In 2005 In 2000 In 1996 Achieved in 2011,2012 Achieved in 2004 Achieved in 2000 Start up in 2013 13.5t/ha

Demonstrations and yield component of super hybrid rice Years/Place Variety Panicles (m2) Plant height (cm) Spikelet Seed set (%) GW (g) Yield (t/hm2) HI 2011/HN Y-1 303.71 134.3 57735.27 90.8 26.47 13.9 0.55 2012/HN Y-8188 259.47 119.3 52975.13 96.7 26.85 13.77 0.54 2013/HN Y-900 227.89 128.6 77162.22 93.5 26.7 14.82 0.57 XLY-2 287.86 116.7 62349.63 92.7 26.5 13.58 Y-6 299.4 124.63 68263.27 93 28 14.41 0.56 23 23

The number of super hybrid rice certified by MOA (2005~2013) SHR Varieties released & certificated The number of super hybrid rice certified by MOA (2005~2013) year 2005 2006 2007 2008 2009 2010 2011 2012 2013 total 3-line 20 7 3 / 4 6 5 52 2-line 2 1 21 22 12 9 73 By 2013, 73 rice hybrids were certified as “super rice”. Among them: 52 three-line hybrid rice varieties, 21 two-line hybrid rice varieties,

3) HR seed production ★ Its difficult to utilize heterosis in rice Dr. J.N.Rutger －Earlirose male-sterile plants averaged 2.4% open-pollinated seed-set From《Innovative approaches to rice breeding》 ★ The key problem for the HR commercialization ● lower pollen load ● Panicle is not exerted out easily ● Out-crossing rate is very low

The technology of Hybrid seed production Optimized ◎　The average yield in China 274.5kg/ha in 1975 1.66 t/ha in 1985 2.25 t/ha in 1990 ◎　Yield on a large scale 3~4.5 t/ha in 1990s ◎　The highest record yield 7.39 t/ha (0.113 ha) ,Hunan China in 1993 Commercial hybrid rice yield and hybrid rice seed yield in China (1976-2008)

Yield of seed production of HR:2.7-3.0t/hm2 A new way for hybrid rice seed production: mechanized techniques New Model of R:A Pollination by UAV Ratio of R:A 6~8:40~80 hills outcrossing rate: 45% Yield of seed production of HR:2.7-3.0t/hm2 The pollination, GA spraying and the ration of A:R lines by the unmanned aerial vehicle (UAV)

4) Field management of HR Chinese hybrid rice agronomists developed the systematic methods for high-yielding field management: "3-ding”Cultivation Technology for Super Hybrid Rice Rice Precise and Quantitative Cultivation Technique for High Yield The Improved System of Rice Intensification (MSRI) Nitrogen-efficiency and lodging resistance cultivation technique

3. Constraints in HR development The Area of HR at a Standstill for Years (17.6 million ha in 1991) Lack of japonica hybrid rice with strong heterosis (0.1 mil ha, only 1%-2% in China). Great yield gap for super hybrid rice (about 5.16 t/ha). Lack of the breakthrough combinations but HR varieties released yearly with about 400. Need of the breakthrough for the mechanized technology of HR seed production.

4. Main Reasons for the success of HR Strong Govt. Leadership: be committed, supportive, well organized to HR R and D. High efficient organization and coordination: Leading Group, Consultancy Committee, frequent conferences; Resources and techniques: qualified, distributive and shared Technology generation and dissemination: R and D cooperation networks, Training, yield trials, on-farm demonstration , study tour and propaganda 源库流的问题几乎包括了玉米产量形成的全过程 源库流的调节是玉米群体质量调控的重要内容。 源大库足“流”畅是玉米高产的基本要求 在源库流关系上明确产量主要限制因子，对于高产育种和栽培都具有重要指导意义

5. Prospect: HR development in the future A new idea of super high yield breeding proposed by Prof. L.P. Yuan in 2009 plant height(m)

New approach for high yield of HR Grain yield = Harvest Index X Biomass Nowadays the harvest index (HI) is much higher (above 0.5). Further lifting of rice yield ceiling should rely on increasing biomass because further improvement of HI is quite limited. From view point of morphology, Further raising rice yield ceiling should rely on increasing biomass. Increasing plant height is an effective and feasible way to increase biomass

The key scientific ideas of the new strategy Morphological improvement with utilization of the heterosis simutaniously Higher biomass by plant height increased Further utilize heterosis by exploitation of indica/japonica heterosis, favorable genes favorable (QTLs) Maintain high harvest index （>0.5) Enhancing the lodging resistant; Recent yield targets: 15t/ha（i.e., single rice）

Thank you ！ 源库流的问题几乎包括了玉米产量形成的全过程 源库流的调节是玉米群体质量调控的重要内容。 源大库足“流”畅是玉米高产的基本要求 在源库流关系上明确产量主要限制因子，对于高产育种和栽培都具有重要指导意义