Characterization of Fertility Alteration in 8 Environment-sensitive Genic Male-sterile Lines of indica Rice

Eight indica ( Oryza sativa L.) environment-sensitive genic male-sterile (EGMS) lines, 2-2S, K1405S, F131S, 2136S, Pei-Ai 64S, 1290S, GD-IS and N17S, were sequentially seeded with 10-15 d interval at three sites, Wuhan in 1997, Guiyang in 1997 and Sanya in 1997 and 1998, China. The results of investigations on self-sterilities showed that all of eight EGMS lines had stable sterile periods of longer than 30 d at Wuhan. They can be used for seed production of two-line hybrid rice, but can not reproduce themselves. Their stable sterile periods were shorter than 30 d at Guiyang, they can reproduce themselves and can not be used for hybrid seed production. In Sanya, their stable sterile periods were longer than 150 d, all of eight lines can be used for seed production in summer and autumn and reproduce themselves in winter. The fertility of all eight lines were sensitive to temperature. The sensitive stages, sensitive duration and critical point of temperatures (CPT) of fertility alteration in various lines were different. The sensitive stages of 2-2S and K1405S were from 18 d to 9 d before heading, the sensitive durations were 7-10 d and the CPTs were 23.7-24.5 degreesC. The sensitive stage, sensitive duration and CPT of F131S were from 17 to 5 d before heading, 13 d and 24.3-24.7 degreesC, respectively. The sensitive stage, sensitive duration and CPT of 2136S were from 18 to 12 d before heading, 7 d and 24.6-25.1 degreesC:, respectively. The sensitive stages, sensitive durations of Pei-Ai 64S, 1290S, N17S and GD-1S were from 24 to 13 d before heading and 10-13 d. And their CPTs were 24.6-25.1 degreesC, 25.5-26.2 degreesC, 25.4-26.1 degreesC,, and 24.1-24.7 degreesC, respectively.

Development of Isogenic Lines of Morphological Markers in Indica Rice

A total of 28 accessions of rice (Oryza sativa L.) germplasms were collected from International Rice Research Institute (IRRI), Philippine and Gene Pool of China National Rice Research Institute (CNRRI). These germplasms were attributed as a Donor Gene Pool, consisting of 27 diverse morphological marker lines with each having a unique valuable trait (gene), and the recurrent parent-Zhefu 802, an early-season indica cultivar in south of China in the 1980s and 1990s. A set of 27 isogenic lines with 2-3 markers on every chromosome were obtained after more than 10 successive backcrosses. Moreover, 15 accessions of double-marker isogenic lines were developed by further pyramiding markers on the same chromosome. Key agronomic traits such as heading date, plant height, tillering number and panicle length in this set of lines were similar to that of Zhefu 802 except the marker traits.

Correlation between Stigma Characteristics and Outcrossing Rate of Five Photo-thermo Sensitive Genic Male Sterile(PTGMS) Rice Lines

[Objective] This study aimed to investigate the correlation between stigma characteristics and outcrossing rate of photo-thermo sensitive genic male sterile（PTGMS） rice lines. [Method] The stigma exsertion rate, stigma size and stigma vitality of Chuang 5S, Yun 9S, Peiai 64 S, P88 S and Y58 S were analyzed. Using R608 as a male parent, outcrossing rate of these five PTGMS lines was determined. [Result]There were differences in stigma exsertion rate, stigma size and stigma vitality among five PTGMS lines. P88 S exhibited the best stigma characteristics and relatively strong stigma vitality, with the stigma exsertion rate of 95.94%, stigma vitality coefficient of 2.71, outcrossing rate of 84.02% in the field. Peiai 64 s had the minimum stigma length（2.41 mm） and stigma width（1.10 mm） and the weakest stigma vitality, with the stigma exsertion rate of 93.04%, outcrossing rate of 65.47%. Yun 9S,Y58 S and Chuang 5S displayed medium stigma characteristics, among which Yun9 S exhibited higher outcrossing rate than other two PTGMS lines. Correlation analysis showed that stigma exsertion rate, stigma length, stigma width, stigma area and stigma volume were significantly correlated to outcrossing rate（P〈0.05）, with correlation coefficients of 0.897, 0.934, 0.934, 0.939 and 0.937, respectively. However,there was no significant correlation between stigma vitality coefficient and outcrossing rate. [Conclusion] Based on stigma characteristics of five PTGMS lines, auxiliary pollination measures should be adopted reasonably to save production costs and improve hybrid rice seed yield.

Comparative Studies on the Changes of Microtubule Distribution and Reorganization During the Meiotic Stages of Development in Normal (IR36) and a Temperature/photoperiod Sensitive Male Sterile Line (Peiai 64S) of Rice ( Oryza sativa )

Changes in the pattern of organization of microtubules in the meiotic stages of development of pollen (i.e. from pre-meiotic interphase to more or less metaphase I) of a normal (IR36) and a temperature/photoperiod sensitive male sterile line (Peiai 64S) of rice were studied using immunofluorescence confocal microscopy. In IR36, from pre-meiotic interphase to metaphase I, the pattern of microtubule distribution in the meiocytes underwent a series of changes. Some new organizational patterns of microtubules (that have not been described before) were observed during microsporogenesis, including the existence of a broad band of perinuclear microtubules at the diakinesis stage of development. The pattern of microtubule distribution in the meiocytes of the male sterile line, Peiai 64S, was quite different front that seen in IR36. In Peiai 64S, the microtubules showed abnormal patterns of distribution from pre-meiotic interphase to metaphase I. For example the broad band of perinuclear microtubules seen at diakinesis in IR36 was much disorganized and loosened in Peiai 64S. The spindles formed were also very abnormal and different from the normal spindle. The appearance of abnormal microtubule distribution in the early stages of microsporogenesis may contribute to the malformation and ultimate abortion of pollen in Peiai 64S.

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.

Mapping of QTLs Related to Grain Weight Using Chromosome Single Segment Substitution Lines in Rice

Grain weight, one of the major factors determining rice yield, is a typical quantitative trait control ed by multiple genes. With Guangluai 4 as recipient and Nipponbare as donor, a population of 119 chromosome single segment substitution lines had been developed. Correlation analysis between grain weight and grain shape by SPSS revealed that 1 000-grain weight shared extremely significant posi-tive correlation with grain length and length-width ratio, but no significant correlation with grain width and thickness. The QTL analysis of grain weight was carried out using one-way analysis of variance and Dunnett＇s test. Nineteen stable QTLs re-sponsible for grain weight were identified over two years. Al 19 QTLs were identi-fied on al chromosomes except for chromosome 10 and 12 at a significance level of P≤0.001. Among them, 10 QTLs had a positive effect and were derived from the Nipponbare al ele, the additive effect of these QTLs ranged from 0.49 to 2.74 g, and the contributions of the additive effects ranged from 2.00% to 11.05%. Another 9 QTLs had a negative effect and were al derived from Guangluai 4 al ele, the ad-ditive effect of these QTLs ranged from 0.60 to 2.35 g, and the contributions of the additive effects ranged from 2.40% to 9.84%. The results provide a basis for the fine mapping and gene cloning of novel locus associated with rice grain weight.

Study on Seed Vigor of Rice Photo-ThermoSensitive Genic Male Sterile(PTGMS) Line with Low Temperature Storage

To explore the change rule of seed vigor and seed application technology of rice PTGMS lines with low temperature storage,Zhun S,Y58S,Feng S and Guangzhan 63-4S stored in the seed bank with the temperature of 8-10 ℃ and with the relative humidity of 55% for different years were used as the materials to study the properties of seed vigor,growth and development in the field, etc. The re- sults showed that the germination potential, germination rate, germination index and vigor index of rice PTGMS lines seeds showed downward trends as the time of low temperature storage prolonged,and all of the germination rates of the seeds within five years were over 60%. The conductivity in the soaked solution of seeds was significantly increased with the prolonged low temperature storage time. Seedling rate,root length,white root number,base stem width and seedling dry weight of seeds showed downward trends as the time of low temperature storage prolonged. There was no significant difference between two integral seedling qualities of seeds within 4-6 years in low temperature storage. Compared with the germination rate of low temperature storage seeds soaking for 6,12 and 24 h,the appropriate soak time of long time low temperature storage or low vigor seeds was 12 h. A serious natural aging phenomenon occurred in low temperature storage seeds after out of the storage. The germination rate of seeds placed under natural condition for 50 d decreased obviously. The duration from seeding to heading of Zhun S,Y58S and Feng S shortened as the time of low temperature storage prolonged, while the vari- ation of panicle duration of all PTGMS lines was not obvious. In conclusion, rice PTGMS line seeds with low temperature storage in 5 years could still be used nor- mally. However.seeds after low temperature storage should be sowed as soon as possible, and the soaking time of low vigor seeds should be shortened,and the possibility that the decreased duration from seeding to heading should also be con- sidered.

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.

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.

Breeding and Application of WA-type Indica CMS Line Ningxian 2A in Rice

Ningxian 2A is a new Indica CMS line with WA type sterile cytoplasm in rice. It was developed from the testcross between WA type indica CMS line Jin 23A and the single plant selected from the F_4 population of the cross Wuxiang 2B×Jin 23B followed by successive backcross over years. In September, 2014, it was technically identified by Jiangsu Provincial Crop Variety Appraisal Committee. Ningxian 2A has the characteristics of complete pollen sterility, high outcrossing rate, good plant type, high combining ability and good grain quality. Among its hybrids, Ningxian you 8（ Ningxian 2A×R08） was registered and released for commercial production in Jiangsu Province in May, 2015.

Breeding and Application of Indica PTGMS Line Yan 220S in Rice

Yan 220S, which is derived from a natural variant plant discovered among the population of Guangzhan 63S and developed by Jiangsu Agricultural Research Institute for Coastal Regions through pedigree selection over years, is an indica photo- and thermo-sensitive genic male sterile （PTGMS） line in rice. In 2013, it was technically identified by Jiangsu Crop Variety Appraisal Committee. Its hybrid Yan-liangyou 2208 （Yan 220S/Yanhui 888）, a new two-line hybrid rice combination with high yield and good grain quality, was subsequently developed and released for commercial production by the National Crop Variety Appraisal Committee in 2013.

Study on Photosynthetic Characteristics of a Rice CMS Line Renong 1A with Yellow-green Revertible Leaves

[Objective] Understanding the photosynthetic characteristics of the rice CMS line Renong 1A with leaf color markers can promote its utilization in breeding. [Method] The wild type （WT） of rice spontaneous mutant with virescent-yellow leaves that was the male parent of Renong 1A was selected as the control, and the contents of photosynthetic pigments, photosynthetic parameters, light-response curve and chlorophyll fluorescence kinetic parameters of Renong 1A were measured at the tillering stage. [Result] New leaves of Renong 1A yellowed obviously in the early period. Chlorophyll content, photosynthetic rate and values of parameters Pdt, F/Fm Fv＇/Fm＇,φPSII, and ETR of Renong 1A were all significantly lower than those of the control. However, there was no difference between Renong 1A and the control in terms of photosynthetic rate of leaves turning green. [Conclusion] The photosynthetic capacity of new leaves of Renong 1A decreased at the early stage due to the reduction of chlorophyll content, and then it became normal at the later stage with the increase of chlorophyll content.

Study on the combination ability and stability of rice restore line Yihui1577

Yihui1577 was bred after over 10 generations by crossing R16 and Yunan purple rice. Many new hybrid rice varieties have been bred with Yihui1577, of which Gangyou1577. Ilyou1577, Nyou1577 and Yixiang1577 were certified by Sichuan Crop Variety Release Committee, Gangyou1577 and Yixiang1577 were certified by National Crop Variety Release Committee. The series varieties bred with Yihui1577 have been used widely with accumulated area of 1,200,000 ha. By using Yihui1577 as breeding material, an excellent restore line Yihui3003 and some other restore lines, such as Yihui1979 and Yihui315 with good quality and combination ability have been bred. Analysis of combination ability and stability of restore line Yihui1577 was made by using agricultural experimentation and statistical approach. The combination ability analysis was made by not complete dual crossing method with 4 CMS lines and 6 restorer lines. It shows that the general combination ability effect value of Yihui1577 is 0.226, while Minghui63 is -0.113. So the general combination ability of Yihui1577 is prior to that of Minghui63. Stability analysis shows that the F value of Gangyou1577 is 1.81378, while Sanyou63 is 1.40327, both doesn＇t reach significant level. So Gangyou1577 has the same stability as Sanyou63.The same conclusion is got in the stability analysis of the breeds of II you 1577, Yixiang1577, Nyou 1577 and so on.

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.

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.

Flowering Characteristics of Japonica Rice Male Sterile Lines Adaptive to High Temperature and Summer Drought

[Objective] This study was conducted to screen japonica rice male sterile lines with good flowering characteristics under high temperature and summer drought. [Method] The flowering habits of 23 japonica male sterile lines, such as flowering peak, single-panicle flowering duration, single-plant flowering duration, en- closed glume rate, glume-opening angle, the distance between glumes, stigma exer- tion rate and sigma vitality, were compared with II-32A as a control under above weather condition. [Result] The flowering characteristics had significant difference a- mong the tested rice lines. Compared with the control, three japonica rice male ster- ile lines 35478A, 35489A and 35502A had better flowering traits, such as earlier flowering peak, longer single-panicle flowering duration, longer single-plant flowering duration, lower rate of enclosed glumes, larger glume angle and larger distance be- tween glumes. 35478A performed its flowering peak from 10：00-11：00 am, 1.5 h earlier than the control; 35489A exhibited higher stigma exertion rate, up to 76.23%; and 35502A had higher stigma vitality, up to 84.68%, which kept longer time than that of the CK. [Conclusion] The results will provide high-quality resources for the breeding of new japonica rice varieties with high-yield and high-temperature toler- ance.

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.

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.

Effect of High Temperature on Outcrossing Characteristics at the Fertility Sensitive Period of Photo-thermo Sensitive Genic Male Sterile (PTGMS) Rice Lines

Four sterile lines （Peiai64S, Y58S, Guangzhan 63-2S and H638S） and the restoring line R527 were used as materials. Five temperature gradients （24, 27, 30, 33 and 36 ℃ in artificial climate chamber） and the natural temperature （as control） were treated to the four sterile lines for 6 d in the fertility sensitive period of heading to flowering stage, respectively, to study the effects of temperature on physiological biochemical characteristics of young panicles and outcrossing characteristics. The results showed that the percentages of exerted stigma of Peiai 64S and Y58S were the highest at 27 ℃, which were 86.81% and 86.06%, respectively, while the percentages of exerted stigma of Guangzhan 63-2S and H638S were the highest at 24 ℃, which were 76.24% and 81.76%, respectively; the stigma viability of Peiai 64S, Y58S, Guangzhan 63-2S and H638S were the best at 24 ℃, which were 1.96, 2.12, 1.74 and 1.94, respectively; the outcrossing rates of Peiai 64S, Guangzhan 63-2S and H638S were the highest at 24 ℃, which were 58.87%, 54.22% and 50.50%, respectively, while the outcrossing rate of Y58S was the highest at 27 ℃, which was 58.96%; and the contents of peroxidase （POD） of the four sterile lines increased significantly at 33 ℃ compared with the control, while the contents of malondialdehyde （MDA） and proline increased significantly at 36 ℃ compared with the control. There were differences in temperature sensitivity between the male sterile lines, and the 24 ℃ treatment during the sensitive period was the best for the fertility sensitive period of Peiai 64S, while 27 ℃ was the best temperature for Y58S, Gangzhan 63-2S and H638S.

QTLs Mapping for Salinity Tolerance at Seedling Stage or Rice(Oryza sativa L.) Using Chromosome Segment Substitution Lines

In this study, a population of chromosome segment substitution lines （CSSLs） derived from the cross between 9311 （indica） and Nipponbare （japonica） was employed to map the quantitative trait loci （QTLs） for salt tolerance under the salt stress simulated with 0.5% NaCI, using survival rate as the index. The data were analyzed by QTL IciMapping v3.1, and the results showed that one QTL （QSsr3） related to salt tolerance was located in the vicinity of the marker RM1350 on chromosome 3, into a genetic interval of 113.2-132.8 cM, with a contribution rate of 17.75%. The additive effect was 10.9, indicating that the QTL derived from the parent Nipponbare improved the salt tolerance of rice at seedling stage. This study will provide a theoretical basis for the selection of salt tolerant rice germplasm.