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.

Study on High-Eefficient Breeding Technology Platform of Dwarf Male-Sterile Wheat

[Purpose] The paper is to discuss the high-efficient breeding technology platform of dwarf male-sterile wheat.[Method] Conclusion was made upon the development of the creation and application of dwarf male-sterile wheat,and the conception of introducing haploid breeding technology into the technological system of high-efficient breeding of dwarf male-sterile wheat was proposed.[Result] Dwarf male-sterile wheat not only retains the properties of total male abortion of Taigu Genetic Sterile Wheat,steady sterility,and high hybrid rate after open pollination,but also has the characteristic of high dwarfing ability of Aibian No.1,becoming a comparatively ideal population modification means.At present,the application of dwarf male-sterile wheat mainly includes germplasm resources platform of population modification by using dwarf male-sterile wheat,technology platform of constructing new high-efficient breeding system by using dwarf male-sterile wheat,and production platform of using dwarf male-sterile wheat to breed new varieties.Through the introduction of haploid breeding into the already established wheat high-efficient breeding system,a new system of high-efficient biological breeding technology of dwarf male-sterile wheat was constructed theoretically.[Conclusion] The study provides references to the further study and application of dwarf male-sterile wheat.

Characterization of Reverse Thermo-sensitive Genic Male Sterile Lines in Cold Regions of China

Yannong S, a reverse thermo-sensitive genic male sterile （TGMS） line, is sterile at high temperature and fertile at low temperature. In the present study, the fertility of Yannong S and the japonica reverse TGMS lines derived from it was in- vestigated in Harbin, Heilongjiang from 2013 to 2015. The results showed that Yan- nong S was completely male sterile in July and August （the hottest months of the year in Harbin）, and the seed setting rate of bagged panicles was 0. We also in- ferred that the critical temperature for inducing the sterility of Yannong S was higher than 28 ℃. Most of the japonica reverse TGMS lines derived from Yannong S were completely aborted in July and August, with a seed setting rate of 0. It can be con- cluded that the reverse TGMS lines derived from Yannong S are genetically stable lines that have a long period of sterility in cold regions, so they may be the ideal genetic materials for the development and in-situ seed production of japonica hybrid rice.

Electron-Microscopic Study of Microsporogencsis in Male-sterile and Male-fertile Grain Amaranth(Amaranthus hypochondriacus L.)

A comparative study of microsporogenesis in male-sterile and male-fertile grain amaranth was conducted using electron-microscopy.The onset of microsporogenesis breakdown in male-sterile is at the mononuclear pollen stage following the release of microspores from tetrads.Abnormality in the behavior of degenerated tapetum was observed, which failed to envelope individual microspores after their release, leading to an abnormal vacuolation in the mononuclear pollen grains.As a result, the normal thickening of pollen wall could not occur and pollen grains could not engorge, causing male sterility.

Research Advances in Molecular Mechanism of Thermo-sensitive Genic Male Sterility in Plants

Thermo-sensitive genic male sterile （TGMS） lines have specific superiority in heterosis utilization of crops. So far, thermo-sensitive genic male sterile lines have been found in many plants and are widely used in two-line hybrid breeding. With the rapid development of molecular biology, the molecular nature of thermo-sensitive genic male sterility has been revealed, which lays the foundation for further devel- opment and utilization of thermo-sensitive genic male sterile lines. In this study, the molecular mechanisms of fertility conversion of plant thermo-sensitive genic male sterile lines were reviewed from gene molecular mapping and gene differential ex- pression, and the mechanisms of gene differential expression in thermo-sensitive genic male sterile lines were further discussed.

Brassinosteroids mediate the effect of high temperature during anthesis on the pistil activity of photo-thermosensitive genetic male-sterile rice lines

Brassinosteroids(BRs)play critical roles in a wide range of plant developmental processes.However,it is unknown whether and how BRs mediate the effect of high temperature(HT)stress during anthesis on the pistil activity of photo-thermosensitive genetic male-sterile(PTSGMS)rice(Oryza sativa L.)lines.This study investigated the question.Three pot-grown PTSGMS rice lines were subjected to HT stress during anthesis.The contents of 24-epibrassinolide(24-EBL)and 28-homobrassinolide(28-HBL),the major forms of BR in rice plants,and levels of reactive oxygen species(ROS)or antioxidants(AOS),hydrogen peroxide(H2O2),1-aminocylopropane-1-carboxylic acid(ACC),ascorbic acid(AsA),and catalase activity in pistils,were determined.HT stress significantly reduced the contents of both 24-EBL and 28-EBL relative to those under normal temperatures,but the reduction varied by PTSGMS line.A line with higher BR contents under HT stress showed lower contents of ACC and H2O2,higher catalase activity and AsA content in pistils,and higher fertilization rate,seed-setting rate,and seed yield when the line was crossed with a restorer line,indicating that higher levels of BRs increase HT stress resistance.Applying 24-EBL,28-HBL or an inhibitor of BR biosynthesis confirmed the roles of BRs in response to HT stress.The results suggest that BRs mediate the effect of HT stress on pistil activity during anthesis and alleviate the harm of HT stress by increasing AOS and suppressing ROS generation.

Thermo-sensitive polymer nanospheres as a smart plugging agent for shale gas drilling operations

Emulsifier-free poly（methyl methacrylate-styrene） [P（MMA-St）] nanospheres with an average particle size of 100 nm were synthesized in an isopropyl alcoholwater medium by a solvothermal method. Then, through radical graft copolymerization of thermo-sensitive mono- mer N-isopropylacrylamide （NIPAm） and hydrophilic monomer acrylic acid （AA） onto the surface of P（MMA- St） nanospheres at 80 ℃, a series of thermo-sensitive polymer nanospheres, named SD-SEAL with different lower critical solution temperatures （LCST）, were prepared by adjusting the mole ratio of NIPAm to AA. The products were characterized by Fourier transform infrared spectroscopy, transmission electron microscopy, thermogravimetric analysis, particle size distribution, and specific surface area analysis. The temperature-sensitive behavior was studied by light transmittance tests, while the sealing performance was investigated by pressure transmission tests with Lungmachi Formation shales. The experimental results showed that the synthesized nanoparticles are sensitive to temperature and had apparent LCST values which increased with an increase in hydrophilic monomer AA. When the temperature was higher than its LCST value, SD- SEAL played a dual role of physical plugging and chemical inhibition, slowed down pressure transmission, and reduced shale permeability remarkably. The plugged layer of shale was changed to being hydrophobic, which greatly improved the shale stability

Genetic Analysis and Preliminary Mapping of a Highly Male-Sterile Gene in Foxtail Millet(Setaria italica L.Beauv.) Using SSR Markers

Breeding of male-sterile lines has become the mainstream for the heterosis utilization in foxtail millet,but the genetic basis of most male-sterile lines used for the hybrid is still an area to be elucidated.In this study,a highly male-sterile line Gao146A was investigated.Genetic analysis indicated that the highly male-sterile phenotype was controlled by a single recessive gene a single recessive gene.Using F 2 population derived from cross Gao146A/K103,one gene controlling the highly male- sterility,tentatively named as ms1,which linked to SSR marker b234 with genetic distance of 16.7 cM,was mapped on the chromosome VI.These results not only laid the foundation for fine mapping of this highly male-sterile gene,but also helped to accelerate the improvement of highly male-sterile lines by using molecular marker assisted breeding method.

Plant Temperature and Its Simulation Model of Thermo-Sensitive Genic Male Sterile Rice

Plant temperature （Tp） and its relations to the microclimate of rice colony and irrigation water were studied using a thermo-sensitive genic male sterile （TGMS） rice line, Pei＇ai 64S. Significant differences in the daily change of temperature were detected between Tp and air temperature at the height of 150 cm （TA）. From 8：00 to 20：00, Tp was lower than TA, but they were similar during 21：00 to next 7：00. The maximum Tp occurred one hour earlier than the maximum TA, though they both reached the minimum at 6：00. Tp fluctuated less than TA. At the same height, during 6：00-13：00, Tp was higher than air temperature （Ta）, and Tp reached the maximum one hour earlier than Ta. During the rest time on sunny day, Tp was close to or even a little lower than Ta. On overcast day, Tp was higher than Ta in the whole day, and both maximized at the same time. In addition, Tp was regulated by solar radiation, cloudage and wind speed in daytime, and by irrigation water at night. The present study indicated that a TA of 29.6℃ was the critical point, at which Tp was increased or decreased by irrigation water. Tp and the difference between water and air temperatures showed a conic relation. Tp fluctuation was also regulated by the absorption or reflection of solar radiation by leaves during daytime and release of heat energy during nighttime. By analysis on correlation and regression simulation, two models of Tp were established.

Mechanism of Sterility and Breeding Strategies for Photoperiod/Thermo-Sensitive Genic Male Sterile Rice

To understand the male sterility mechanism of photoperiod/thermo-sensitive genic male sterile [P（T）GMS] lines in rice, the research progress on genetics of photoperiod and/or temperature sensitive genic male sterility in rice was reviewed. A new idea was proposed to explain the sterility mechanism of P（T）GMS rice. The fertility transition from sterile to fertile is the result of cooperative regulation of major-effect sterile genes with photoperiod and/or temperature sensitive genes, but not the so-called pgms gene in P（T）GMS rice. The minor-effect genes, which exhibit accumulative effect on sterility, are the important factors for the critical temperature of sterility transition. The more minor-effect genes the sterile line holds, the lower the critical temperature of sterility transition is. The critical temperature of sterility transition will be invariable if all the minor-effect genes are homozygous. The strategies for breeding P（T）GMS rice were also proposed. The selective indices of critical photoperiod and temperature for sterility transition should be set according to varietal type and ecological region. Imposing selection pressure is a key technology for breeding P（T）GMS rice with lower critical temperature for sterility, and improving the comprehensive performance of the whole traits and combining ability is vital for breeding P（T）GMS rice lines.

Molecular Markers and Candidate Genes for Thermo-Sensitive Genic Male Sterile in Rice

The discovery of thermo-sensitive genic male sterility(TGMS) has led to development of a simple and highly efficient two-line breeding system. In this study, genetic analysis was conducted using three F_2 populations derived from crosses between IR68301 S, an indica TGMS rice line, and IR14632(tropical japonica), Supanburi 91062(indica) and IR67966-188-2-2-1(tropical japonica), respectively.Approximately 1:3 ratio between sterile and normal pollen of F_2 plants from the three populations revealed that TGMS is controlled by a single recessive gene. Bulked segregant analysis using simple sequence repeat(SSR) and insertion-deletion(InDel) markers were used to identify markers linked to the tms gene. The linkage analysis based on the three populations indicated that the tms locus was located on chromosome 2 covering the same area. Using IR68301S × IR14632 F_2 population, the results showed that the tms locus was located between SSR marker RM12676 and InDel marker 2gAP0050058. The genetic distance from the tms gene to these two flanking markers were 1.10 and 0.82 cM, respectively.InDel marker 2gAP004045 located between these two markers showed complete co-segregation with the TGMS phenotype. In addition, InDel marker vf0206114052 showed 2.94 cM linked to the tms gene using F_2 populations of IR68301S × Supanburi 91062. These markers are useful tool for developing new TGMS lines by marker-assisted selection. There were ten genes located between the two flanking markers RM12676 and 2gAP0050058. Using quantitative real-time PCR for expression analysis, 7 of the 10 genes showed expression in panicles, and response to temperatures. These genes could be the candidate gene controlling TGMS in IR68301S.

Characterizations and identification of the candidate gene of rice thermo-sensitive genic male sterile gene tms5 by mapping

Previous study indicated that the thermo-sensitive genic malesterile(TGMS) gene in rice was regulated by temperature.TGMS rice plays an important role in hybrid rice production,because the application of the TGMS system in two-line breeding is laborsaving,timesaving,simple,inexpensive,efficient,and eliminating the limitations of the cytoplasmic male sterility(CMS) system.'AnnongS' is the first discovered and deeply studied TGMS rice lines in China.'AnnongS-1' and 'Y58S',two derivatives of TGMS line AnnongS,were both controlled by a single recessive gene named tms5,which was genetically mapped on chromosome 2.In this study,three populations('AnnongS-1' × 'Nanjing11','Y58S' × 'Q611',and 'Y58S' × 'Guanghui122') were developed and used for the molecular fine mapping of the tms5 gene.By analyzing recombination events in the sterile individuals using a total of 125 probes covering the tms5 region,the tms5 gene was physically mapped to a 19-kb DNA fragment between two markers 4039-1 and 4039-2,which were located on the BAC clone AP004039.After the construction of the physical map between two markers 4039-1 and 4039-2,a member(ONAC023) of the NAC(NAM-ATAF-CUC-related) gene family was identified as the candidate gene of the tms5 gene.

Characterization and fine mapping of RTMS10, a semi-dominant reverse thermo-sensitive genic male sterile locus in rice

The discovery and application of environment-sensitive genic male sterile(EGMS) rice germplasm provide an easy method for hybrid rice breeding and have made great contributions to hybrid rice production. Typically, the photoperiod-and thermosensitive GMS(P/TGMS) lines utilized in two-line hybrid systems are male sterile under long day or/and high temperature but fertile under short day or/and low temperature conditions. However, Yannong S(Yn S), a reverse TGMS(rTGMS) line, is sterile under low temperature(<29℃) and fertile under high temperature(>29.5℃). Here, we report a genetic study on the rTGMS trait in Yn S. Interestingly, the F1 plants of the cross between Yn S and a cultivar, L422, were male sterile at 22℃ and completely fertile at 27℃. Moreover, the segregation ratio of fertile and sterile individuals in Yn S/L422 F2 populations changed from 1:3.05 to 2.95:1 when the ambient temperature increased, showing that the rTGMS trait exhibits semidominance in Yn S. We further found a locus on chromosome 10, termed RTMS10, which controls the rTGMS trait in Yn S. We then finely mapped RTMS10 to a ~68 kb interval between markers ID13116 and ID1318 by Yn S/L422 BC6 F2 populations. A near iso-genic line(NIL) NL1 from the BC6 F3 generation was developed and the pollen of NL1 became abnormal from the meiosis stage under low temperature. In summary, we identified an rTGMS locus, RTMS10, and provided co-segregated markers, which could help to accelerate molecular breeding of rTGMS lines and better understand the rTGMS trait in rice.

Marker-Assisted Breeding of Thermo-Sensitive Genic Male Sterile Line 1892S for Disease Resistance and Submergence Tolerance

Rice line 1892S is an elite thermo-sensitive genic male sterile(TGMS)line for two-line hybrid rice production.However,1892S is susceptible to rice blast,bacterial blight and submergence.Here we reported the introduction of blast resistance(R)gene Pi9,bacterial blight R gene Xa21 and submergence tolerance gene Sub1A into 1892S genetic background through backcrossing and marker-assisted selection.The improved TGMS line 31892S and its hybrids conferred disease resistance to rice blast and bacterial blight,and showed submergence tolerance for over 14 d without significant loss of viability.The sterility-fertility conversion of 31892S was similar to that of 1892S.31892S and its derived hybrid rice had similar agronomic traits and grain quality with 1892S and the control hybrid rice,respectively.The newly developed 31892S provided an improved TGMS line for two-line hybrid rice production with disease resistance to rice blast and bacterial blight,and submergence tolerance with no yield penalty or change in grain quality.

Genetic Analysis and Mapping of a Thermo-sensitive White Stripe-Leaf Mutant at Seedling Stage in Rice(Oryza sativa)

A thermo-sensitive white stripe-leaf mutant （tws） was selected from the M2 progeny of a japonica variety, Jiahua 1, treated by ^60 Co γ-radiation. In comparison with the wild type parent, the mutant displayed a phenotype of white stripe on the 3rd and 4th leaves, but began to turn normal green on the 5th leaf when grown at low temperatures （20℃ and 24℃）. Furthermore, the content of total chlorophyll showed an obvious decrease in the leaves with white stripe. These results suggest that the expression of the mutant trait was thermo-sensitive and correlated with the leaf age of seedlings. The genetic analysis indicated that the mutant trait was controlled by a single recessive nuclear gene, designated as tws. In addition, by using SSR markers and an F2 segregating population derived from the cross between the tws mutant and 9311, tws was mapped between the markers MM3907 and MM3928 with a physical distance of 86 kb on dce chromosome 4.

An easily recoverable thermo-sensitive polyelectrolyte as draw agent for forward osmosis process

As a potential solution to the crises of energy and resources, forward osmosis(FO) has been limited by the development of draw agents. An ideal draw agent should be able to generate high osmotic pressure and can be easily recovered. In this study, a thermo-sensitive polyelectrolyte of poly(N-isopropylacrylamide-co-acrylic acid)(PNA)is developed as an efficient draw agent, and two easy and simple methods are proposed to effectively recover the polyelectrolytes. After adjusting the pH value of polyelectrolyte solutions to around 6.0, the polyelectrolyte can generate relatively high osmotic pressure, and induce average water fluxes of 2.09 and 2.95 L·m^(-2)·h^(-1) during12 h FO processes when the polyelectrolyte concentrations are 0.20 and 0.38 g·ml^(-1) respectively. After acidifying and heating to 70 °C, the PNA-10 polyelectrolyte can aggregate together because of hydrophobic association and separate from water, so it can be easily recovered by either simple centrifugation or gravitational sedimentation. The recovery ratios of PNA-10 polyelectrolyte in both methods are as high as 89%, and the recovered polyelectrolytes can be reused with almost the same FO performance as fresh ones. The results in this study provide valuable guidance for designing efficient and easily recoverable draw agents for FO processes.

Improving Blast Resistance of a Thermo-Sensitive Genic Male Sterile Rice Line GD-8S by Molecular Marker-Assisted Selection

The broad-spectrum blast resistance gene Pi-1, from donor line BL122, was introduced into a thermo-sensitive genic male sterile rice line GD-8S, which possessed good grain quality but high susceptibility to rice blast, by using backcross breeding and molecular marker-assisted selection. Five elite improved male sterile lines, RGD8S-1, RGD8S-2, RGD8S-3, RGD8S-4 and RGD8S-5, were selected based on the results of molecular marker analysis, spikelet sterility, recovery rate of genetic background and agronomic traits. Thirty-three representative blast isolates collected from Guangdong Province, China were used to inoculate the improved lines and the original line GD-8S artificially. The resistance frequencies of the improved lines ranged from 76.47% to 100%, much higher than that of the original line GD-8S （9.09%）. On the agronomic characters, there were no significant differences between the improved lines and GD-8S except for flag leaf length and panicle number per plant. The improved lines could be used for breeding hybrid rice with high blast resistance.

Differences in Nuclear DNA Between Male-Sterile and Male-Fertile Lines of Sorghum bicolor

Cytoplasmic male sterility(cms)is determined by nuclear-cytoplasmic interactions. Up to now, most studies are focused on the comparison of cytoplasmic DNAs of male-sterile lines and male-fertile lines, and analysis of nuclear DNA has not been documented yet. In order to find out the possible difference in nuclear genome of male-sterile line A1 Tx623 and corresponding male-fertile line Tx623 of sorghum, random amplified polymorphic DNA(RAPD)approach was used to analyze their cytoplasmic and nuclear genomes. Total DNAs of them were amplified at first to screen primers, which were able to generate reproducible bands specific to male-sterile line or male-fertile line. Then the selected primers were used to amplify their mitochon-drial DNA(mtDNA)and chloroplast DNA(cpDNA). The origins of all the polymorphic fragments were analyzed. After ruling out those amplified from cytoplasmic DNA, seventeen polymorphic fragments were determined to be amplified from nuclear DNA. These fragments originated from nuclear DNA indicate that differences in sequence exist between the nuclear DNA of male-sterile line and male-fertile line of sorghum, which do not agree with the traditional standpoint that they have identical nucleus.

A bull-heading water control technique of thermo-sensitive temporary plugging agent

Aimed at the disadvantages of secondary damage to oil layers caused by the conventional bull-heading water control technique, a thermo-sensitive temporary plugging agent for water control was synthesized by water solution polymerization and applied in the field with a new secondary temporary plugging technique. The optimization and performance evaluation of thermo-sensitive temporary plugging agent were carried out through laboratory experiments. The optimized formula is as follows:(6%-8%) acrylamide +(0.08%-0.12%) ammonium persulfate +(1.5%-2.0%) sepiolite +(0.5%-0.8%) polyethylene glycol diacrylate. The thermo-sensitive temporary plugging agent is suitable for formation temperatures of 70-90 ?C, it has high temporary plugging strength(5-40 k Pa), controllable degradation time(1-15 d), the apparent viscosity after degradation of less than 100 m Pa?S and the permeability recovery value of simulated cores of more than 95%. Based on the research results, secondary temporary plugging technique was used in a horizontal well in the Jidong Oilfield. After treatment, the well saw a drop of water cut to 27%, and now it has a water cut of 67%, its daily increased oil production was 4.8 t, and the cumulative oil increment was 750 t, demonstrating that the technique worked well in controlling water production and increasing oil production.

Utilization of recessive male-sterile lines in soybean

Three male-sterile soybean (MS) groups were selected out through various methods. A number of filial generations and hybrid strains with special characters were obtained by the methods of isolated plot and backcross. Among them Jinping 1 and Jinping 2, two sister lines,with higher production,big and pretty seed,green coat and a rich perfume,are suitable to a vegetable in city. It was proved that the MS have wide utilization potential as follows; (a)the MS could solve a lot of troubles of artificial cross ;for instance,the green seed coat can be used as a gene marker; (b)the MS showed a high hybrid vigor and its potential ratio ranged from 132% to 347% ;and (c)the effects of recurrent selection of MS were obvious and the characters of yield of F5 in 18 strains were higher than that of F3.