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.

Effects of Nitrogen Quantity on Grain-filling Characters of Two-line Hybrid Rice with Large Ear

[Objective] The aim was to research effects of N quantity on grain-filling characters of two-line hybrid rice cultivars with large ears. [Method] Peiza 67 and 88, two-line hybridized rice with large ears, were made use of to study on effects of N fertilizer in different quantities （LN： 90 kg/hm2;MN： 180 kg/hm2;HN： 270 kg/hm2） on plumpness and grain-filling characters. [Result] When N fertilizers were excessive, for inferior grains, grain-filling rate decreased and grain-filling time extended, resulting in plumpness decline after degradation of leaves＇ function. When N fertilizers were inadequate, maximal and average grain-filling rates decreased and the differences between superior and inferior grains in grain-filling rate increased, leading to decline of grain＇s weight and plumpness degree. On the other hand, quantity of N fertilizers had little effect on superior grains in plumpness. [Conclusion] The research provided references for reasonable use of N fertilizer and improvement of rice yield and N use.

Breeding of National Authorized Two-line Hybrid Rice Yanliangyou 2208

The breeding processes, characteristics and comprehensive characters of two-line hybrid rice Yanliangyou 2208 and its parents （Yan 220S and Yanhui 888） taking part in the national rice regional test of southern rice regions were introduced. Yanliangyou 2208 was approved by the National Crop Variety Appraisal Committee in 2013. The achievement has been transformed successfully and has been applied in production. It has good development and promotion prospects.

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.

Inheritance Pattern of Two-line Hybrid Rice Resistance to False Smut

[Objective] The aim of this study was to explore the inheritance pattern of the resistance of two-line hybrid rice to false smut. [Method] A moderate susceptible TGMS line 33S as the female parent, was crossed with 30 susceptible restorer lines and 23 resistant restorer lines. Then false smut resistance of the hybrid F1 and parents were identified. [Result] All F1 of 33S crossed with 30 susceptible re- storer lines were susceptible to rice false smut. The F1 of 33S crossed with 23 re- sistant restorer lines showed dominant, incompletely dominant or recessive inheri- tance. Dominant and incompletely dominant inheritance accounted for 87.0%. The diseased panicle rate, disease index, average diseased grains per panicle, and the highest diseased grains per panicle of F1 shared highly significant positive correla- tions with corresponding indicators of restorer lines. [Conclusion] This study provides references for genetic improvement of two-line hybrid rice.

Analysis on Genetic Differences between Parents of Two-line Medium Indica Hybrid Rice by SSR Markers

The genetic diversity of 50 parents or improved lines of two-line medium Indica hybrid rice were analyzed based on 48 pairs of SSR markers. The results showed that the clustering results were highly accordant with the parental pedigree information of the tested materials. The male sterile lines and the restoring lines were clustered into 2 distinct groups （Ⅰ and Ⅱ）, respectively. There were no significant difference in intra-group average genetic distances （GD） between the 2 groups. Most of the male sterile lines from Hunan Longping High-tech Seed Science Research Institute Co., Ltd. were grouped into a single subgroup （Ⅰ-4） under the male sterile group with the largest average GD compared with the rest male sterile subgroups. Huazhan derived lines and Guangzhou inbred lines were founded to be independent of 9311 and its derivatives or closely-related lines. Meanwhile, they have obviously been differentiated into 2 subgroups （Ⅱ-2 and Ⅱ-3）. The lines derived from Huazhan （or genetic closely related lines） showed a genetic homogenization phenomenon. Therefore, the Huazhan-derived lines should be further improved by the utilization of germplasms with good quality, high resistance and wide adaptability from Southern China to widen the GD between Huazhan derived lines and the male sterile subgroup developed by Hunan Longping High-tech Seed Science Research Institute Co., Ltd., so as to further promote the heterosis level between the groups.

Effect of Different Fertilization Methods on Grain Yield and Nitrogen Agronomy Efficiency of TwoLine Super Hybrid Rice

ln order to explore the effect of broadcast application of urea （BR-U） and surface concentrated-fertilization on grain yield and nitrogen agronomy efficiency of two-line super hybrid rice, this experiment was conducted with super hybrid rice Y-Liangyou 1 and Liangyou 0293 to determine til ering dynamics, SPAD, LAl and dry matter accumulation. lts total N application was 180 kg/hm2 and different application rate and topdressing methods were set respectively. The results showed that grain yield and nitrogen agronomy efficiency were significantly improved with the T3 and T4 nitrogenous fertilizer application model. The improving of grain yield and nitrogen agronomy efficiency was benefited from rational postponing N application. However, neither single application of base fertilizer （T2） nor single application of topdressing fertilizer （T5 and T6） can coordinate the relationship between source and sink. Both grain yield and nitrogen agronomy efficiency were insignificantly under the broadcast application of urea and surface concentrated fertilization.

Performance and High-Yield Cultural Techniques of Two-Line Hybrid Rice Xinlingyou998 at Direct Seeding Cultivation

Xinliangyou 998 is a new indica two-line hybrid rice variety using Xin＇er Sx H0998 as parent. The variety was introduced and demonstrated in Yingshan County in 2016, and showed intermediate growth duration, strong tillering ability, firm culm, tolerance to high fertility and resistance to lodging, tolerance to high tempera- ture and wide range of adaptation, etc. The high-yield cultural techniques were con- cluded.

Breeding of a Water-saving Drought-resistant Two-line Hybrid Rice Variety Wanliangyou 1008

Drought is one of the important factors limiting rice production,and the application of new water-saving rice varieties is urgently needed in rice production.Wanliangyou 1008 bred by Institute of Rice Research,Anhui Academy of Agricultural Sciences is a new two line hybrid rice variety with some good traits,including high drought resistance,high yield and good disease resistance.Wanlingyou 1008 was obtained by crossing drought-resistant high-quality restorer line DJ1008 and disease-resistant two-line sterile line Wan 25S.In this paper,the breeding process,characteristics and seed production technology of the variety were described in detail.

Effects of Tiller-inhibitor on Growth and Yield Formation of Super Early Hybrid Rice Jinyou458

[Objective]The paper was to study the effect of tiller-inhibitor on the growth and yield formation of super early rice Jinyou 458.[Method] Two treatments including spraying tiller-inhibitor（TI） and spraying water（CK） at the SN-n stage were set in the test,the effect of tiller-inhibitor on yield and its components,leaf and plant morphology,field microclimate at booting stage and quality characteristics of rice population were studied.[Result] Spraying tiller-inhibitor could effectively reduce the occurrence of invalid and inefficient tillers,increase the proportion of high effective tillers（tiller with 4 or more leaves） in tiller composition at the maximal tiller stage.The panicle length,spikelets per panicle,spikelets density,number of secondary branches and the secondary spikelets,seed setting rate of rice plant sprayed with tiller-inhibitor were significantly higher than CK.Meanwhile,spraying tiller-inhibitor could increase plant height,biomass weight and leaf area index of rice population during middle and late stages of rice growing,improve the leaf temperature of top three leaves and the light transmittance of rice population at the booting stage,increase the leaf SPAD value,thus enhance photosynthetic capacity of rice plants.It also showed the characteristics of elongating the internode of rice plant and decreasing the stem thickness,etc.[Conclusion] The panicle-bearing rate of stems and tillers,effective panicles,spikelets per panicle,seed setting rate and 1 000-grain weight had coordinately increased after spraying tiller-inhibitor,thus increased the yield.

Effect of Microbial Inoculum Fertilizer on the Growth and Yield of Super Hybrid Rice

The research designed four application quantities of 0, 2 250, 4 500 and 6 750 g/hm2 based on Liangyou No. 959 to explore the effects of microbial inocu- lure fertilizer on growth and yield of super hybrid rice. The results indicate that the treatment with traditional fertilizers and 4 500 g/hm2 HYT microbial inoculum fertilizer performed the best, where average number of tiller was 2.4 more than that of the control; the number of effective ear was 11.05% higher; yield increased by 10.3%. These have demonstrated that microbial inoculum fertilizer is of great value in pro- motion on super hybrid rice production.

Stem Morphological Structure of Super Hybrid Rice and Its Relationship with Lodging Resistance

The stem morphology and anatomical structure of rice, as well as their relationship with lodging resistance, were studied with six super hybrid rice varieties as the tested materials. The results showed that the rice varieties with stronger lodging resistance were characterized by shorter basal elongated internodes and thicker stem base. The lodging index of hybrid rice was positively correlated with the lengths of the first, second and third basal elongated internodes （P〈0.01） and was negatively correlated with the thickness of basal stem wall and the number and area of small vascular bundles, the area of large vascular bundles and the total area of vascular bundles of the second basal elongated internode （P〈0.05）. The correlations between lodging index and plant height, maximum culm diameter of stem base and minimum culm diameter of stem base were not significant. It indi- cates that the lodging resistance of super hybrid rice can be improved by shorten- ing the length of basal elongated internodes, thickening the wall of stem base and increasing the number and area of vascular bundles of basal elongated internodes.

Prospects for Study on High-yielding Cultivation Techniques of Super Hybrid Rice

Great progresses have been made in super hybrid rice in China. From the perspectives of dry matter production, nutrient absorption, sink and source, pho-tosynthesis, graln fiI ing and roots, the eco-physioIogical characteristics and high-yielding cuItivation techniques of super hybrid rice in China were discussed. In addi-tion, the probIems that restricted the high and stabIe yielding of super hybrid rice were analyzed, and the deveIopment directions of high-yielding cuItivation techniques for super hybrid rice were also discussed.

Effects of Nitrogen Rate on Blast Resistance and Grain Yield of Super Early Hybrid Rice Luliangyou 996

Super early hybrid rice Luliangyou 996 was the materials for this experiment. This paper focused on the influences of different nitrogen rates on blast resistance and grain yield. Studies suggested that with the increase of nitrogen rate,there were no distinct changes in the sick grain rate and sickness index, but the fifth grade of panicle blast rate rose gradually. When the nitrogen was too much（270 kg/hm^2）, the sick grain rate and sickness index were the highest. The yield rose along with the addition of nitrogen if the amount of applied nitrogen was within the range of 0 to 180 kg/hm^2, and the yield would decrease with the addition of nitrogen if the amount of applied nitrogen was over 180 kg/hm^2. Considering the sickness, output and economic benefit of rice, it is better to applied 90 to 135 kg/hm^2 of nitrogen for Luliangyou 996.

Changes in the Activities of C4 Pathway Enzymes and Stable Carbon Isotope Discrimination in Flag Leaves of Super High-yield Hybrid Rice

Activities of several key enzymes of C-4 photosynthesis pathway and stable carbon isotope discrimination were investigated in flag leaves of a super high-yield hybrid rice (Oryza sativa L.) cv. Peiai 64S/E32 and a traditional hybrid rice cv. Shanyou 63 at different developing stages. Results show that the activity of PEP carboxylase (PEPCase) increased with age of flag leave; the activity of NADP-malate dehydrogenase (NADP-MDH) increased and reached to a peak value at grain filling stage (68-75 d after transplanting), then fell down; the activity of NADP-MDH in cv. Peiai 64S/E32 was much higher than that in cv. Shanyou 63. Before ripening stage (95 d after transplanting), NADP-malic enzyme activity rose gradually. The level of stable carbon isotope discrimination (Delta(13)C) in flag leaves and grains at different developing stages were similar and exhibited a comparative high value at ripening stage. The average Delta(13)C in leaf of cv. Peiai 64S/E32 during different developing stages was 0.43parts per thousand more than that in cv. Shanyou 63.

Relationship Between Grain Yield and Yield Components in Super Hybrid Rice

Chinese super hybrid rice breeding project has developed many new varieties with great yield potential. It is controversial which yield component should be emphasized in super hybrid rice production. The present study was conducted to compare super hybrid rice with common hybrid and super inbred rice and analyze contributions of yield components to grain yield of super hybrid rice under experimental conditions, and evaluate relationships between grain yield and yield components of super hybrid rice in farmer’s paddy fields. Field experiments were done in Changsha, Guidong, and Nanxian, Hunan Province, China, from 2007 to 2009. Eight super hybrid varieties, one common hybrid variety, and one super inbred variety were grown in each location and year. Rice production investigation was undertaken in high-yielding （Guidong）, moderate-yielding （Nanxian）, and low-yielding （Ningxiang） regions of Hunan Province, China, in 2009. Grain yield and yield components were measured in both the field experiments and rice production investigation. Super hybrid rice varieties outyielded common hybrid and super inbred varieties across three locations and years. Yield potential has been increased by 11.4% in super hybrid rice varieties compared with common and super inbred varieties. The higher yield of super hybrid varieties was attributed to improvement in panicle size. Panicles per m2 had the highest positive contribution to grain yield with the exception under yield level of 10.0 to 12.0 t ha-1, and was positively related to grain yield in farmer’s field at all of the high-, moderate-, and low-yielding regions. Our study suggests that panicle per m2 ought to be emphasized in super hybrid rice production.

Practices and Prospects of Super Hybrid Rice Breeding

The great progress in super rice breeding both in China and other countries has been made in recent years. However, there were three main problems in super rice breeding： 1） the super rice varieties were still rare; 2） most super rice varieties exhibited narrow adaptability; and 3） current breeding theories emphasized too much on the rice growth model, but they were unpractical in guidance for rice breeding. According to the authors＇ experience on the rice breeding, the breeding strategies including three steps （super parent breeding, super hybrid rice breeding and super hybrid rice seed production） were proposed, and the objectives of each step and the key technologies to achieve the goals were elucidated in detail. The super parent of hybrid rice should exhibit excellent performance in all agronomic traits, with the yield or sink capacity reached the level of the hybrid rice control in regional trials. The super hybrid rice combination should meet the following criteria： good rice quality, wide adaptation, lodging resistance, resistance to main insects and diseases, and the yield exceeded above 8% over the control varieties in the national and provincial regional trials. To achieve the goal, the technical strategies, such as selecting optimal combination of the parents, increasing selection pressure, paying more attention to harmony of ideal plant type, excellent physiological traits and all the agronomic traits, should be emphasized. The yield of seed production should reach 3.75 t/ha and 5.25 t/ha for the super hybrid rice combinations derived from early-season and middle-season types of male sterile lines, respectively. The main technologies for raising seed production yield included selecting optimum seed production site, using the male sterile line with large sink capacity and good outcrossing characteristics, and improving the amount of the pollen by intensive cultivation of the male parent. According to the technologies of the three-step breeding on super hybrid rice, two super rice parents, including a male parent 996 and a thermo（photo）-genic male sterile [T（P）GMS] line C815S, were bred. Furthermore, a super early hybrid rice combination, Luliangyou 996, which could be used as a double-season early rice variety in middle and lower reaches of the Yangtze River, China, was bred by using the super rice varlet3, 996 as the male parent, and several hybrid rice combinations with higher yield than control variety in regional trials both of Hunan Province and state were bred with the T（P）GMS line C815S as the female parent.

Yield potential and stability in super hybrid rice and its production strategies

China＇s Super Hybrid Rice Breeding Program has made significant progress over the past two decades. In this paper, we reviewed our studies on the yield potential and stability in super hybrid rice and discussed the strategies for super hybrid rice production. The results of our studies show that rice yield potential has been increased by 12% in super hybrid cultivars as compared with ordinary hybrid and inbred cultivars. The higher grain yields in super hybrid rice cultivars are attributed to larger panicle size coupled with higher biomass production or higher harvest index. However, grain yields in super hybrid rice cultivars vary widely among locations depending on soil and climatic factors. Therefore, it is important to tailor target yield to local conditions in super hybrid rice production. The target yield for super hybrid rice production can be determined by the average yield method or the regression model method. Improving soil quality is critical to achieving the target yield in super hybrid rice production. Favorable crop rotations such as rice-oilseed rape and novel soil management practices, such as biochar addition, are effective approaches to improve soil quality. It is needed to develop simplified cultivation tech- nologies for super hybrid rice to meet the changes in socioeconomic environments during the period of transition. There are such technologies as no-tillage direct seeding and mechanized transplanting at high hill density with single seedling per hill.