Solutions to Insecurity Problems in Seed Production of Two-line Hybrid Rice

The insecure problems of seed production have seriously hampered the healthy and sustainable development of two-line hybrid rice.The safety issues on seed production of two-line hybrid rice and current research situation were pointed out in this paper.The three main reasons for unsafety in seed production of twoline hybrid rice were unsuitable site selection,high critical sterility-inducing temperature and the drift of critical temperature.In this paper,strategies and measures were put forward based on many years of practice.It could minimize the risk in seed production of two-line hybrid rice by selecting dual-purpose genic male sterile line with lower critical sterility-inducing temperature and long lower temperature resistant time.Based on the climate data and climatic demands of the ＂three safe-periods＂ in seed production,a new idea for determining appropriate bases and periods for seed production was proposed by using computer technology,which solved the aimless selection of sites and periods for the seed production of two-line hybrid rice.Besides,we established a system of single plant selection and identification method and original seed propagation with cyclic cold water,which could reduce the generation number of original seed propagation in seed production and avoid the drift of critical sterility-inducing temperature.This paper improved the seed production safety in the three aspects of seed nature,seed source and seed production site.

Analysis of Heterosis, Combining Ability and Heritability of Cadmium Content in Brown Rice of Three-line Indica Hybrid Rice

Five cytoplasmic male sterile （CMS） lines were used as parents in an incomplete diallet cross and six restorer lines of rice design （Nc II design）. Thirty hybrid combinations with the same growth period were selected as experimental ma- terials to investigate the heterosis, combined ability and heredity of Cd content in brown rice of indica hybrid rice. According to the results, Cd content in brown rice showed a significantly negative heterosis; the general combining ability and specific combination ability of Cd content in CMS and restorer lines both reached extremely significant level （P〈0.01）, indicating that both genetic improvement of parents and e- valuation of combinations are important to the breeding of hybrid combinations with low accumulation of Cd; the broad-sense heritabitity and narrow-sense heritability of Cd content were both relatively high with slight differences, which respectively reached 97.73% and 80.10%, indicating that Cd content in brown rice mainly de- pends on the additive action of genes; in addition, parent improvement showed bet- ter effect on the selection of early generation.

High-yielding Seed Production Techniques of Disease-resistant and High-quality Hybrid Rice Sterile Line Rong 18A

Rong 18A, bred by Crop Research Institute, Chengdu Academy of Agri- culture and Forestry Sciences is a new blast-highly resistant and high-combining ability hybrid rice sterile line. It is currently one of the male sterile lines in Sichuan Province which combine well high grain quality and rice blast resistance. More than 20 combinations of Rong 18A have passed the national and provincial variety iden- tification. Combined with the climate conditions in Chengdu, the key points of high- yielding seed production techniques for Rong18A are analyzed and summarized in this paper.

Effect of Enhanced UV-B Radiation on Seed Setting Rate and 1 000-grain Weight of Indica Hybrid Rice Restorer Lines

[Objective] This study was to investigate the effect of enhanced UV-B radiation on seed setting rate and 1 000-grain weight of hybrid rice combinations. [Method] The seed setting rate and 1 000-grain weight of 10 new sterile indica restorer lines planted in pots under enhanced UV-B radiation and fluorescent lamps （control） were respectively measured, and the differences were compared. [Result] The enhanced UV-B radiation significantly reduced the seed setting rate of indica restorer lines, and the differences between that UV-B radiation treatment and control all reached extremely significant level. In addition, the enhanced UV-B radiation reduced the 1 000-grain weight of most indica restorer lines, and compared with that of control the difference achieved significant or very significant level. However, the effect of enhanced UV-B radiation on seed setting rate and 1 000-grain weight differed to different indica restorer lines, and the differences among restorer lines tested were significant or very significant, which indicated the possibility to screen antiUV-B radiation rice materials and combinations. Finally, the indica restorer lines 09R-14, Luhui 37 and 10R-7703 which were strongly resistant to UV-B radiation were screened out. [Conclusion] This study laid foundation for breeding hybrid rice varieties resistance to UV-B radiation.

Thoughts and Practice on Some Problems about Research and Application of Two-Line Hybrid Rice

The main problems about research and application of two-line hybrid rice were reviewed, including the confusing nomenclature and male sterile lines classification, the unclear characteristics of photoperiod and temperature responses and the unsuitable site selection for male sterile line and hybrid dce seed production. In order to efficiently and accurately use dual-purpose genic male sterile lines, four types, including PTGMS （photo-thermo-sensitive genic male sterile rice）, TGMS （thermo-sensitive genic male sterile rice）, reverse PTGMS and reverse TGMS, were proposed. A new idea for explaining the mechanism of sterility in dual-purpose hybrid rice was proposed. The transition from sterile to fertile was involved in the cooperative regulation of major-effect sterile genes and photoperiod and/or temperature sensitive ones. The minor-effect genes with accumulative effect on sterility were important factors that affected the critical temperature of sterility transfer. In order to make better use of dual-purpose lines, the characterization of responses to photoperiod and temperature of PTGMS should be made and the identification method for the characterization of photoperiod and temperature responses of PTGMS should also be put forward. The optimal ecological site for seed production could be determined according to the historical climate data and the requirements for the meteorological conditions during the different periods of seed production.

Research on Theories and Techniques of Irrigation for Safeguarding Seed Production of Two-Line Hybrid Rice

By inducing frequency, intensity and duration of lower temperature in the middle and last ten-day periods in August in the rice-growing areas of southern China, increasing temperature for safe seed production was defined as 2℃. During the sensitive period of fertility, characters of panicle height and canopy structure of TGMS rice, Pei＇ai64S, were measured. Results showed that temperature changes caused by irrigation in fields were below 40 cm of rice plant, and heating effect was significant at 20 cm when the temperature was increased by 3.1 ℃. For the present study, the following irrigation techniques were put forth： the water depth of 15-20 cm, current water used, irrigating after 17：00 and bailing at 10：00 in sunny or cloudy weather, irrigating on whole day, in shady or rainy weather, increasing inflows and outflows in large fields. In the present experiment, pollen fertility and self-fertilized seed setting rate accepted that the techniques were feasible and effective for against lower temperature and safeguarding seed production of two-line hybrid rice.

Causes of Poor Grain Plumpness of Two-Line Hybrids and Their Relationships to the Contents of Hormones in the Rice Grain

This study was designed to elucidate the grain filling characteristics and the causes of poor grain plumpness of some two-line hybrid rice combinations and their hormonal mechanism. Six varieties, including three two-line hybrid rice combinations, that show differences in seed-setting and grain filling, were used. And the contents of starch, sucrose, zeatin （Z） ＋ zeatin riboside （ZR）, indole-3-acetic acid （IAA）, and abscisic acid （ABA）, the ethylene evolution rate, activities of sucrose synthase （SuSase） and starch synthase （StSase） in grains, the seed-setting and grain filling rate were investigated. The correlations amongst these were analyzed. The results showed that the poor grain filling of two-line hybrids was mainly attributed to the higher unfilled grain rate and the lower filling degree of inferior grains. During the early and mid grain filling periods, the sucrose content in inferior grains was greater than that in superior grains for the combinations with poor grain filling, indicating that the substrate concentration was not the principal factor for their slow grain filling and poor grain plumpness of the inferior grains of two-line hybrids. Z ＋ ZR, IAA, and ABA in superior grains were obviously greater than those in inferior grains at early grain filling stage. The maximum and mean contents of Z ＋ ZR, IAA, and ABA were positively very significantly correlated with the maximum and mean grain-filling rate, filling degree, and grain weight. The evolution rate of ethylene was greater in inferior grains than in superior grains and greater for the combinations with poor grain plumpness than those with good grain plumpness at the early or mid filling stages. The evolution rate of ethylene was negatively and significantly correlated with the grain filling rate, the grain filling degree, and the grain weight. Spraying ethephon （ethylene-releasing agent） at the early grain filling stage increased the evolution rate of ethylene, reduced the ABA content and activities of SuSase and StSase, and decreased the grain filling degree and the grain weight. The results were reversed when cobatous nitrate （an inhibitor of ethylene synthesis） was applied. The results suggested that the hormones and their balance play a role in the regulation of grain filling and enzymatic activities, and the poor grain filling is attributed to the low contents of Z ＋ ZR, IAA, and ABA, and the high evolution rate of ethylene in the inferior grains of some two-line hybrid rice combinations. The results suggested that hormones play important roles in the grain filling of some two-line hybrid rice combinations, and their filling degree can be improved by regulating the hormonal contents.

New Temperature Sensitive Genic Male Sterile Lines with Better Outcrossing Ability for Production of Two-Line Hybrid Rice

An investigation was carried out with three newly developed temperature sensitive genic male sterile （TGMS） lines for their floral traits, seed production potential and outcrossing ability in ten cross combinations. In the TGMS lines, fertile pollens had an average diameter of 0.89 mm while the sterile pollens was with 0.02 mm diameter.TS-29-150GY produced the biggest fertile pollens with 0.92 mm and other two lines produced relatively smaller pollens with 0.91 and 0.85 ram. Pollen fertility during the fertility reversion period was an average of 60.7%. TS-29-150GY had the maximum of 66.9% spikelet fertility whereas other two lines （TNAU18S and TNAU60S） had relatively lower spikelet fertility of 27.8% and 26.7%, respectively. Average of 17.00 g of seed yield was obtained in the TGMS lines during the fertility reversion period. TS-29- 150GY had the highest value of 21.20 g of seed yield while TNAU18S and TNAU60S produced 16.6 g and 13.2 g of seed yield, respectively. The low seed production ability of these three TGMS lines was attributed only to the environmental conditions prevailing during the period. All three TGMS lines had considerable outcrossing potential of 41.2%, 24.6% and 25.0%, respectively. The cross combinations viz. TNAU18S/IET21508 （36 g/plant）, TNAU18S/IET21044 （13 g/plant）, TNAU18S/IET21009 （26.5 g/plant）, TNAU60S/CB-09-106 （26.2 g/plant）, TNAU60S/IET21009 （14 g/plant） and TS29-150-GY/DRR 3306 （39.2 g/plant） showed perfect synchronization with acceptable hybrid seed yield, indicating suitability of TGMS system under Indian condition. Based on the outcrossing related traits viz. panicle exertion, angle of glume opening, stigma length and pollen size, TNAU18S was identified as the best, followed by TS-29-150GY.

Utilizing Dominant Early Maturity Genes of Sterile Line UP-3s in Hybrid Rice Breeding to Avoid High Temperature Season

A new sterile line UP-3s, which carries the Dominant Early Maturity Gene (DEMG), was bred on the farm of University of Arkansas at Pine Bluff (UAPB). UP-3s and two check sterile lines, Jin23-A and Xie-A which do not carry the Dominant Early Maturity Gene, were crossed with a group of different maturity restorer lines, PB-1R, PB-5R,PB11, PB-13R, PB-20, PB-21, PB-22R, and PB-23R. Eighteen new hybrid rice combinations of these crosses were then tested at UAPB in 2012 and 2013. The results showed that panicle differentiation (PD) of hybrids from female parent UP-3s (DEMG) crossed with the 8 male parents, were earlier than the hybrids from female parent Jin23-A or Xie-A crossed with the 8 male parents. The PD of these earlier hybrids was before Jun 25 and heading was before July 20. Early PD and heading avoided the high temperature (over 34&degC) period which usually occurs after July 20 in Arkansas. The yields of these earlier maturity hybrids with female parent UP-3s were higher than those of the late maturity hybrids thatwereF1 progeny of sterile lines Jin23-A or Xie-A (these two female parent checks with non-DEMG). These results showed that the DEMG sterile line UP-3s can be adopted in making crosses with later maturity restorer lines to obtain earlier maturity hybrids to avoid the high temperature period in Arkansas.

Studies on the Developmental Genetics of Tiller Number in Three-line Indica Hybrid Rice

Following NCI design, the developmental genetic behavior of tiller number (TN) in three-line indica hybrid rice was studied using additive-dominance developmental genetic models and the corresponding statistical methods. The results showed that dominance effects were predominant for TN. The expression of those additive effects were affected by environment and genotype interaction, but the expression of dominance effects were not affected. Heterosis was the strongest in the middle developmental periods of TN. Additive effects and dominance effects were selectively expressed throughout in the entire tillering developmental stage. Analysis of genetic correlation between TN at different stages and the productive panicles indicated that a close correlation appeared earlier in the populations with higher heterosis than in those with less heterosis. Utilization of heterosis at the middle tillering stage might enhance the final biomass but reduce the percentage of productive panicles.

Influence of Genetic Drift of Restoring Gene (Rf) on Seed Purity of Yuetai A, a Honglian-Type Cytoplasmic Male Sterile Line in Hybrid Rice

The seed samples of Yuetai A, a Honglian （HL） type cytoplasmic male sterile （CMS） line in hybrid rice were investigated to assess the seed purity and to analyze the cause of off-type plants by imitating the biological contaminant to Yuetai A in Nanjing, Jiangsu Province and Lingshui, Hainan Province during 2004-2006. The seed impurity of Yuetai A mainly resulted from the genetic drift of restoring fertility gene （Rt） after biological contamination but not from its sterility unstability. All of the ten maintainer lines, five restorer lines and three thermo-sensitive genic male sterile lines used in the study could pollinate Yuetai A and Yuetai B to produce F1 plants, directly or indirectly resulting in Rf-gene drifting into Yuetai A and generating ＇iso-cytoplasm restoring-generations＇. Furthermore, high outcrossing rate and similar heading date of Yuetai A with many varieties used in rice production might easily result in the biological contamination. After removing all plants with Rf-gene mixed in Yuetai A and preventing Rf-gene drifting into Yuetai A, the seed purity of Yuetai A and Yuetai B had been raised to 100%.

Study on Agronomic Characters and Storage Durability of a New Line of Endurable Storage Hybrid Rice Chucangyou No.1

[Objective] The aim of this study was to investigate the storage durability of a new line of endurable storage hybrid rice Chucangyou No.1 selected by endurable storage genes of Yunhui 290.[Method] Agronomic characters of the new line and the control II you 838 and Shanyou 63 were comparatively studied,and storage durability of the new line and indica,japonica or hybrid rice were also comparatively analyzed by the accelerated ageing test.[Result] Agronomic characters of the new line were excellent,and germination rate in the artificial accelerated ageing test was significantly higher than those in other materials.[Conclusion] Chucangyou No.1 has the better storage durability,which can be used in large scale of extention.

Combining Ability Analysis on Yield Traits of Recombinant Inbred Lines in Hybrid Rice

A population of 140 recombinant inbred lines at F8 generation were obtained after seven successive generations of self-pollination using single seed descent（SSD） method from the F2 hybrids of three-line restorers Luhui 8258 with high combining ability and Yanghui 34. Then, the 140 inbred lines obtained above and their parents Luhui 8258 and Yanghui 34 were crossed with three different types of cyto-plasmic male sterile（CMS） lines（Gang 46 A, Ⅱ-32 A and Lu 98A） according to NCⅡ design. The resulting 426 combinations were planted at Deyang and Suining bases to test the combining ability and inheritance of five yield traits： yield per plant, panicle number per plant, filled grain number per panicle, seed setting rate and 1 000-grain weight. The results showed that the variances of both general and specific combining abilities of the five traits all reached a significant or extremely significant level at the two sites. The broad and narrow heritability of the yield traits（except 1 000-grain weight whose broad and narrow heritability were both over70%） were all below 50% at the two experimental bases, suggesting that the four traits were easily subjected to environment influence. Very significant positive correlation of general combining ability was found between yield per plant and other traits except 1 000-grain weight. The general combining ability variance showed a normal distribution among the recombinant inbred lines at two sites, right in line with inheritance of quantitative traits. So, the genes controlling rice general combining ability can be targeted by QTL mapping.

Breeding and Application of Indica PTGMS Line Yan 161S and Its Hybrid Yanliangyou 1618 in Rice

In 2006, the indica photoperiod- and thermo-sensitive genic male sterile （PTGMS） line Yan 161S was successful y bred by crossing the Guangzhan 63S （♀） and the medium indica breeding material Yan 229 （♂）, which was bred by the Institute of Agricultural Sciences in Coastal Region of Jiangsu Province, after years＇ and generations’ selection in different areas. It passed the identification by Jiangsu Provincial Crop Variety Appraisal Committee in 2013. Crossing the Yan 161S with Yanhui 888, a new high-yielding and high-quality two-line hybrid rice combination Yanliangyou 1618 was developed, and it passed the variety identification in 2014. This paper introduced the breeding processes and characteristics of Yan 161S and Yanliangyou 1618 and the cultivation and seed production techniques of Yanliangyou 1618.

Characterization and evaluation of rice blast resistance of Chinese indica hybrid rice parental lines

The development of resistant varieties and hybrid combinations has been the most effective and economical strategy to control blast disease caused by Magnaporthe oryzae.However, the distribution of major R genes and blast resistance characterization in hybrid rice parents has not been well investigated, resulting in their limited use in hybrid rice blast-resistance breeding. In the present study, 88 elite indica hybrid rice parental lines were evaluated with 30 isolates of M. oryzae collected from the main planting area of indica hybrid rice in China and were characterized for the presence of 11 major resistance genes using molecular markers. The pathogenicity assays showed that four types of hybrid rice parent line showed some resistance to M. oryzae. However, the proportions of highly resistant lines and the mean resistance frequency(RF) varied among the four types, with resistance in decreasing order shown by three-line restorer lines, three-line maintainer lines, two-line sterile lines, and two-line restorer lines. All 88 hybrid rice parental lines carried more than one R gene, but none carried the R genes Pi1 and Pi2. Although Pid3 and Pi9 were present only in three-line restorer lines and Pigm only in three-line maintainer lines, the remaining six R genes(Pib, Pid2, Pi5, Pia, Pi54, and Pita) were present in the four types of hybrid rice parent with significantly different distribution frequencies. The correlation between R genes and resistance reactions was investigated. The results are expected to provide useful information for rational utilization of major R genes in hybrid rice breeding programs.

Progress on Transferring Elite Genes from Non-AA Genome Wild Rice into Oryza sativa through Interspecific Hybridization

The progress of research on transferring elite genes from non-AA genome wild rice into Oryza sativa through interspecific hybridization are in three respects, that is, breeding monosomic alien addition lines （MAALs）, constructing introgression lines （ILs） and analyzing the heredity of the characters and mapping the related genes. There are serious reproductive barriers, mainly incrossability and hybrid sterility, in the interspecific hybridization of O. sativa with non-AA genome wild rice. These are the ＇bottleneck＇ for transferring elite genes from wild rice to O. sativa. Combining traditional crossing method with biotechnique is a reliable way to overcome the reproductive barriers and to improve the utilizing efficiency of non-AA genome wild rice.

Innovation and development of the third-generation hybrid rice technology

The breeding and large-scale application of hybrid rice contribute significantly to the food supply worldwide.Currently,hybrid seed production uses cytoplasmic male sterile(CMS)lines or photoperiod/thermo-sensitive genic male sterile(PTGMS)lines as female parent.Despite huge successes,both systems have intrinsic problems.CMS systems are mainly restricted by the narrow restorer resources that make it difficult to breed superior hybrids,while PTGMS systems are limited by conditional sterility of the male sterile lines that makes the propagation of both PTGMS seeds and hybrid seeds vulnerable to unpredictable climate changes.Recessive nuclear male sterile(NMS)lines insensitive to environmental conditions are widely distributed and are ideal for hybrid rice breeding and production,but the lack of effective ways to propagate the pure NMS lines in a large scale renders it impossible to use them for hybrid rice production.The development of"the third-generation hybrid rice technology"enables efficient propagation of the pure NMS lines in commercial scale.This paper discusses the establishment of"the thirdgeneration hybrid rice technology"and further innovations.This new technology breaks the limitations of CMS and PTGMS systems and will bring a big leap forward in hybrid rice production.

Practice and Thought on Developing Hybrid Rice for Super High Yield by Exploiting Inter-subspecific Heterosis

Since the breakthrough of grain yield owing to the development of dwarf rice and three-line system hybrid rice, rice breeding for high yield hardly had showed significant progress in the next successive two decades. It was considered that utilizing heterosis between subspecific varieties (Oryza sativa L.) would be an effective approach to increase yield further. During 1987-1993, an indica-japonica hybrid Yayou 2 yielded as high as 10.5 t/ha; however, it failed to be commercialized because of seed purity problem due to non-uniform emasculation by chemical agent in seed production, and sensitivity of seed setting in Ft plants to environmental conditions. In the past decade, two inter-subspeific hybrids, Liangyoupeijiu (Peiai 64S/9311, javanica/indica) and Liangyou E32 (Peiai 64S/E32, javanica/ laponica); both of them exhibited grain yield higher than 10.5 t/ha, and were widely judged as the pioneers of super hybrid rice. Liangyoupeijiu has been successfully popularized over 4 million hectare in wide climatic areas, while Liangyou E32 made a yield record and offered a model of plant ideotype for super hybrid rice. It was considered that in combination with plant ideotype, active physiological functions, and wide-range adaptability to ecological conditions, exploitation of indica-japonica heterosis would be the key approach for super hybrid rice breeding.

Resistance Evaluation of Some Chinese Leading Rice Maintainer, Restorer lines and Their Hybrids to Magnaporthe grisea

Six isolates of Magnaporthe grisea were selected to inoculate on 10 Chinese leading maintainer lines (B-lines), 14 restorer lines (R-lines) and their F1 hybrid plants. In the tested rice materials, R-lines were proved to be more resistant to blast than B-lines. The resistance frequency of about 25% F1 hybrid plants was less than their parents. In addition, 26 isolates of M. grisea collected from different rice growing areas of China were inoculated on 13 new improved hybrid rice combinations. The resistance frequencies of 5 improved hybrids were better than those of the controls and leading varieties in rice production of China.

Breeding Technology of eui-hybrids of Rice

Induced mutants of elongated uppermost internode (eui) were directly obtained after irradiation on maintainer (B) and restorer (R) lines of hybrid rice. CMS lines(A) with eui (referred to as eA), restorer lines with eui (referred to as eR) as well as hybrid rice with eui gene (referred to as e-hybrids)were developed. The results showed that the eui mutation frequency averaged 0.10‰ at a range of 0.02‰- 0.47‰ varying with applied dosage and mutated materials. Genetic diversities of eui stock, and a new eui gene, temporarily designated as eui2(t), was identified. eA lines have a significantly elongated uppermost internode causing better panicle exsertion and possessing higher outcrossing potential, e-hybrids increased plant height by 3 -10 cm and headed 2-3 days earlier, as well as showed greater grain weight resulting in higher yield potential than that of corresponding original hybrids. The technology systems without GA3, a few of using GA3 in hybrid rice seed production, and the establishment of e-hybrid rice breeding system were also discussed in this paper.