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.

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.

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.

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.

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.

Breeding and Application Techniques of a New Rice Variety Liliang-you 3822 with High Yield and Disease Resistance

Liliangyou 3822 is a novel indica hybrid rice variety that exhibits disease resistance,high yield,lodging resistance,and late maturity.It employs a self-selected two-line sterile line,Li 38S,and a self-selected restorer line,R22.This variety was subjected to a regional test of indica late-maturing groups in the middle and lower reaches of the Yangtze River in 2020.The results demonstrated that the average yield of the variety was 9.95 t/hm 2,which was 10.67%higher than that of the control Fengliangyou 4,indicating a highly significant yield increase.In the continuous test in 2021,the average yield was 9.74 t/hm 2,representing a 6.52%increase over the control,which also exhibited a significant increase.Finally,the average yield of the two years regional test was 9.84 t/hm 2,which was 8.58%higher than that of the control.In the 2021 production test,the average yield of the variety was 9.32 t/hm 2,which was 12.19%higher than that of the control,indicating a remarkably significant yield increase.In 2022,the variety was validated by the National Crop Variety Approval Committee(GSD 20220143).

Thinking on the Successful Promotion of Twoline Hybrid Rice C Liangyou 34

C Liangyou 343 is a two-line hybrid rice combination with multiple spikes, which has high yield and wide adaptability, and with superior rice quality to other combination. Analysis showed that innovation was the key to the successful promotion of C Uangyou 343, and if it could be applied to the dry-cultivation successfully, a new market could be opened up.

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.

High-yielding Seed Production Techniques and Characteristics of Hybrid Rice Yanliangyou 888

According to seed production of Yanliangyou 888, a two-line hybrid rice variety, the research reviewed seed production characters, as follows： to improve outcrossing rate and spike size provided sufficient spikes of female/male parents. The seed production techniques include scientific arrangement of seeding interval, cultivating strong multi-tiller seedlings, scientific arrangement of row ratio of inter- cropping with adequate basic seedlings, scientific water and fertilizer management, improving outcrossing rate by synthesis measures, disease/insect control, and rein- forcing production management.

Development of Hybrid Rice Variety FY7206 with Blast Resistance Gene Pid3 and Cold Tolerance Gene Ctb1

Hybrid rice Fanyou 7206（FY7206）, derived from the cross between a sterile line Fanyuan A and a restorer line Fuhui 7206, was bred by the Rice Research Institute, Fujian Academy of Agricultural Sciences, China. FY7206 was characterized by moderate blast resistance, cold tolerance, as well as wide adaptability, and high yields. The blast resistance results indicated that the frequencies of blast races in race B, race C and the total resistance frequency for FY7206 were 95.5%, 100.0% and 97.2%, respectively. The disease resistance results showed that the leaf blast grade for FY7206 was level 1 and panicle blast was level 5. The indoor spray results indicated that FY7206 was resistant to 11 isolates of Magnorpathe oryzae. The blast resistance of FY7206 might be derived from the high expression of blast resistance gene Pid3. The results for simulated cold resistance in an artificial climate chamber indicated that the cold tolerance for FY7206 was moderate at the booting and flowering stages. The cold tolerance results also indicated that FY7206 could be tolerant to temperatures as low as 10 °C at the seedling stage. The q RT-PCR results showed that the expression of cold tolerance gene Ctb1 in FY7206 was relatively high. These results suggested that FY7206 is a hybrid indica rice variety with good comprehensive characteristics, including blast resistance and cold tolerance.

SAR-3——a resource of diploid true breeding hybrid rice

In 1988, we found strain 9003 from compositehybridization of indica and japonica. 9003 pos-sesses characteristics of twin seedlings withfrequency of 15-20% of the investigated popu-lation. The highest frequency was 45.68%.Afterwards, we discovered that some twinseedlings had variabilities in chromosome set.As to a pair of twin plants, it may be 2n-1n,2n-3n, 3n-3n or 1n-1n. The changes in chro-mosome set also occurred in the plants devel-oped from mono-embryo seeds. All of thesevariants nearly account for 1% in our con-trolled experiment. We named the triploidplants SAR-3 and found that SAR-3 could be aresource of diploid true breeding hybrid rice.

Effects of a GA3/4PU-30 compound (90-09) on leaf senescence of hybrid rice

90-09 is a cmpound made with GA3,4PU-30and some trace elements(B ete.).To studythe effects of 90-09 on the leaf senescence ofhybrid rice leaf,the changes of activities ofsome degradation enzyme and the contents ofendogenous hormones(ABA,ZRs,GAs,andIAA),protein,and nucleic acids were mea-sured during leaf senescence. Shanyou 63,an indica hybrid rice,wasused in this experiment.Rice plants were firstsprayed with 90-09(115 ml/hm~2) solution 10d after heading,followed by two more sprayswith once a week.The controlled plants weresprayed with water only.

Improving the resistance of the rice PTGMS line Feng39S by pyramiding blast,bacterial blight,and brown planthopper resistance genes

Knowledge of rice(Oryza sativa L.)genes and various DNA markers can be used in genomic breeding programs aimed at developing improved elite rice cultivars.We used an efficient genomic breeding approach to pyramid four resistance genes(Pi2,Xa23,Bph14,and Bph15)in the popular photoperiod-and thermosensitive genic male sterile(PTGMS)rice line Feng39S.We performed foreground selection for the target genes,followed by recombinant selection and background selection.This process reduced the sizes of the genomic segments harboring the target genes(566.8 kb for Pi2,1143.9 kb for Xa23,774.7 kb for Bph14,and 1574.9 kb for Bph15)and accelerated the recovery of the recurrent parent genome to proportions ranging from 98.77%to 99.16%,thus resulting in four near-isogenic lines.To assemble the four resistance genes in Feng39S,we performed a double-way cross combined with foreground and background selection to generate two improved lines of Feng39S(Pi2+Xa23+Bph14+Bph15)with a recurrent parent genome recovery of 98.98%.The two lines showed agronomic performance,grain quality,and fertility–sterility transition characteristics similar to those of the original Feng39S line.The newly developed PTGMS lines and corresponding hybrid combinations were resistant to various field blast isolates and seven representative isolates of bacterial blight.At the seedling stage,the lines also showed resistance against brown planthopper.This study provides an efficient and accurate genomic breeding approach for introducing desirable traits into PTGMS lines.

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.

Hybrid Rice Research and Development in China and Its New Progress

Hybrid rice's history and its development up to date are described in the paper. Compared with the conventional rice, hybrid rice can increase grain yield by about 20%. Hybrid rice breeding in China has been advancing along the three-line method, to two-line method, to super high-yielding rice application strategically and its success in higher degree will be seen by the utilization of distant heterosis cooperated with biotechnology.