Understanding a key gene for thermosensitive genic male sterility in rice

<正>Recently,a joint research team led by Chuxiong Zhuang of South China Agricultural University and Xiaofeng Cao of Institute of Genetics and Development Biology,Chinese Academy of Sciences published their work on the cloning and molecular characterization of the gene thermosensitive genic male sterile 5(tms5)in rice in Nature Communications[1].This is the result of a long-term collaboration representing an important advance in male sterility research in crops.     

Double-stranded RNA and male sterility in rice

Summary Double-stranded RNA (dsRNA) was isolated from rice Oryza sativa ssp. japonica, but not from other subspecies. The dsRNA has been found in all of the examined cytoplasmic male-sterile (CMS) lines of BT (Chinsurah Boro II)-type rice, but was not detected in their companionate maintainer lines. It is uniquely and positivley correlated with the CMS trait in BT-type rice. Recently, the dsRNA was also found in a nuclear malesterile (NMS) rice, Nongken 58s, but was not found in its normal Nongken 58. The molecular weight of this dsRNA was estimated to be about 18 kb. Electron microscopic analysis reveals that it is linear snapped. The double strandedness of the RNA molecules was characterized by CF-11 cellulose column chromatography and nuclease treatments. It bound to CF-11 cellulose in the presence of 15% ethanol. It was sensitive to RNase A at low salt concentrations, but insensitive to DNase I, SI nuclease, and RNase A at high salt concentrations. The dsRNA was detected in both mitochondrial and cytoplasmic fractions. Dot-blot hybridization reveals that there is no sequence homology between this dsRNA and mtDNA, but there is homology between this dsRNA and nuclear genomic DNA. We have not been able to transmit this dsRNA to fertile rice.     

The causes of genetic male sterility in 3 soybaen lines

Summary The cause of male sterility in 3 soybean lines, TGM 103-1, N-69-2774 and TGM 242-4 was studied. In TGM 103-1, which was both male and female sterile, two different abnormalities were associated with sterility. Precocious movement of a few chromosomes at the metaphase I stage resulted into the production of non-functional pollen while cells which underwent apparent normal meiotic division had disintergration of the tapetal cell wall immediately after the free microspore stage leading to the starvation and subsequent death of the developing microspores. In lines N-69-2774 and TGM 242-4, both of which were partially sterile, male sterility resulted from a failure of cytokinesis after the telophase II stage. Meiosis proceeded normally but the 4 microspores after telophase II failed to separate into pollen grains and degenerated thereafter.     

水稻“三明”显性核不育基因的定位

"三明显性核不育水稻"突变体是由福建省三明市农业科学研究所于2001年在杂交组合"SE21S/Basmati370"的F2代群体中发现的。其不育性受1个显性基因控制(将该基因命名为SMS)。经过多代回交,该显性不育基因已导入籼稻品种佳福占的遗传背景中(将该不育材料称为佳不育)。为了定位SMS,文章将佳不育与粳稻品种日本晴杂交,并将F1与佳福占测交,构建了一个作图群体。利用SSR和INDEL标记,通过混合分离分析和连锁分析,将SMS定位于第8号染色体上两个INDEL标记ZM30和ZM9之间,约99 kb的区间内。该结果为克隆SMS奠定了基础。     

Molecular mapping of a rice gene conditioning thermosensitive genic male sterility using AFLP, RFLP and SSR techniques

The discovery and application of the thermosensitive genic male sterility (TGMS) system has great potential for revolutionizing hybrid seed production technology in rice. Use of the TGMS system in two-line breeding is simple, inexpensive, efficient, and eliminates the limitations associated with the cytoplasmic-genetic male sterility (CMS) system. An F₂ population developed from a cross between a TGMS indica mutant, TGMS–VN1, and a fertile indica line, CH1, was used to identify molecular markers linked to the TGMS gene and to subsequently determine its chromosomal location on the linkage map of rice. Bulk segregant analysis was performed using the AFLP technique. From the survey of 200 AFLP primer combinations, four AFLP markers (E2/M5–600, E3/M16–400, E5/M12–600, and E5/M12–200) linked to the TGMS gene were identified. All the markers were linked to the gene in the coupling phase. All except E2/M5–200 were found to be low-copy sequences. However, the marker E5/M12–600 showed polymorphism in RFLP analysis and was closely linked to the TGMS gene at a distance of 3.3 cM. This marker was subsequently mapped on chromosome 2 using doubled-haploid mapping populations derived from the crosses IR64×Azucena and CT9993×IR62666, available at IRRI, Philippines, and Texas Tech University, respectively. Linkage of microsatellite marker RM27 with the TGMS gene further confirmed its location on chromosome 2. The closest marker, E5/M12–600, was sequenced so that a PCR marker can be developed for the marker-assisted transfer of this gene to different genetic backgrounds. The new TGMS gene is tentatively designated as tms4(t). Received: 13 July 1999 / Accepted: 27 July 1999     

无花粉型水稻温敏核不育系籼S的育性表现与细胞学观察

温敏核不育水稻籼S是从优质常规稻籼黄占自然突变而来的一个无花粉型光温敏核不育种质资源。在广州(23°08′N)自然条件下,一年中具有明显的“可育-不育-可育”的育性转换,5月初至10月底为稳定不育期。在人控光温条件下,低温诱导其由不育转为可育需要较长的持续时间,日均温21℃需7d以上,23.5℃需15d以上。细胞学观察表明其无花粉败育主要是由减数分裂时期的异常引起的,表现为小孢子母细胞粘连与液泡化、减数分裂受阻于前期Ⅰ的细线期、进行无丝分裂与异常的胞质分裂,始终没有正常四分体的形成,而是产生大小不同、核数不等的异常细胞,并最终解体消失。其花粉败育特点不同于以往研究过的光温敏核不育水稻,具有花粉败育时期早而败育彻底的特点。     

Mitochondrial DNA modifications associated with cytoplasmic male sterility in rice

Summary Mitochondrial DNA was isolated from fertile and cytoplasmic male sterile lines of rice. Restriction analysis showed specific modifications in the male sterile cytoplasm. In addition to the major mitochondrial DNA, three small plasmid-like DNA molecules were detected by agarose gel electrophoresis in both cytoplasms. An additional molecule was specifically found in the sterile cytoplasm. These mitochondrial DNA modifications support the hypothesis of the mitochondrial inheritance of the cytoplasmic male sterility in rice.     

大豆细胞核雄性不育基因研究进展

雄性不育是指植物雄蕊不能正常生长和产生有活力花粉粒的现象。利用雄性不育突变体开展杂交育种工作,是快速提高作物单产的有效途径。目前,通过杂种制种已大幅度提高了水稻(Oryza sativa L.)、玉米(Zea mays L.)和小麦(Triticum aestivum L.)等作物的产量。大豆(Glycinemax(L.)Merr.)作为自花授粉作物,通过人工去雄生产杂交种子不仅困难而且经济上不可行。由于适用于杂交种生产的不育系资源短缺,目前大豆还没有实现大规模杂种优势利用。因此,快速实现大豆杂种优势利用迫切需要鉴定稳定的大豆雄性不育系统。本文总结了大豆细胞核雄性不育(genic male sterility, GMS)突变体及不育基因研究进展,同时结合拟南芥(Arabidopsis thaliana)、水稻和玉米中已报道的细胞核雄性不育基因,从反向遗传学的角度,为大豆核雄性不育基因的鉴定提供依据。     

Photoperiod- and thermo-sensitive genic male sterility in rice are caused by a point mutation in a novel noncoding RNA that produces a small RNA

Photoperiod- and thermo-sensitive genic male sterility (PGMS and TGMS) are the core components for hybrid breeding in crops. Hybrid rice based on the two-line system using PGMS and TGMS lines has been successfully developed and applied widely in agriculture. However, the molecular mechanism underlying the control of PGMS and TGMS remains obscure. In this study, we mapped and cloned a major locus, p/tms12-1 (photo- or thermo-sensitive genic male sterility locus on chromosome 12), which confers PGMS in the japonica rice line Nongken 58S (NK58S) and TGMS in the indica rice line Peiai 64S (PA64S, derived from NK58S). A 2.4-kb DNA fragment containing the wild-type allele P/TMS12-1 was able to restore the pollen fertility of NK58S and PA64S plants in genetic complementation. P/TMS12-1 encodes a unique noncoding RNA, which produces a 21-nucleotide small RNA that we named osa-smR5864w. A substitution of C-to-G in p/tms12-1, the only polymorphism relative to P/TMS12-1, is present in the mutant small RNA, namely osa-smR5864m. Furthermore, overexpression of a 375-bp sequence of P/TMS12-1 in transgenic NK58S and PA64S plants also produced osa-smR5864w and restored pollen fertility. The small RNA was expressed preferentially in young panicles, but its expression was not markedly affected by different day lengths or temperatures. Our results reveal that the point mutation in p/tms12-1, which probably leads to a loss-of-function for osa-smR5864m, constitutes a common cause for PGMS and TGMS in the japonica and indica lines, respectively. Our findings thus suggest that this noncoding small RNA gene is an important regulator of male development controlled by cross-talk between the genetic networks and environmental conditions.     

Inheritance of spontaneous male sterility in peas

A plant with a mutant phenotype was observed in a Longittee cultivar. The plant was late in maturity, had white-translucent anthers, and was male sterile. The inheritance of this mutant was studied in a cross involving the mutant and the mother parent and their F₁, F₂, F₃ and BC₁F₁ generations. The results suggested that the sterile character was genetic and due to a recessive gene.     

Fine mapping of a gene for non-pollen type thermosensitive genic male sterility in rice (Oryza sativa L.)

The thermo-sensitive genic male sterility (TGMS) lines play a crucial role in two-line hybrid rice production. For a practical TGMS line, the stability of male sterility is one of the most important technical indicators. In this study, XianS, a spontaneous mutant with stable male sterility from an indica rice cultivar Xianhuangzhan, was classified as a non-pollen type TGMS line. The critical non-pollen sterility point temperature of XianS was determined as 27°C. Genetic analysis demonstrated that the non-pollen sterility in XianS was controlled by a single recessive gene. Using SSR markers and bulked segregant analysis, the TGMS gene in XianS was fine mapped to a 183 kb interval between RMAN81 and RMX21 on chromosome 2. Two markers, 4039-1 and RMX14 completely cosegregated with this gene. Allelism test indicated that the non-pollen phenotype in seven non-pollen type TGMS lines from different sources, XianS, AnnongS-1, Q523S, Q524S, N28S, G421S, and Q527S is caused by the same TGMS gene. Although the location of TGMS gene in XianS is close to the gene OsNAC6, a previously identified candidate gene of tms5 in AnnongS-1, the sequence of OsNAC6 and its promoter region was identical in TGMS line XianS, AnnongS-1, and wild-type Xianhuangzhan. These results suggest that the non-pollen type TGMS trait probably be controlled by the same TGMS gene in different TGMS rice lines, but its real candidate gene still need to be further studied and identified.     

Inheritance of spontaneous male sterility in pigeonpea

 A male-sterile plant was observed in the UPAS-120 cultivar of pigeonpea (Cajanus cajan). The plant was about 5–7 days late-flowering and had white translucent anthers with complete pollen sterility. The inheritance of this spontaneous male sterility was studied in a cross involving the mutant and fertile UPAS-120, including their F₁, F₂, BC₁F₁ and BC₂F₁ generations. The results suggested that the male sterility was genetic and due to a recessive gene. Received: 12 November 1996/Accepted: 17 January 1997     

Gene-dependent male sterility and plastomes in Oenothera

The plastids in the cells of the tapetum in anther of Oenothera are involved in the development of male sterility (mst). We combined nuclear homozygosity for each of the two mst genes with the four different plastomes of Oenothera and demonstrated that in both cases the sterile anther phenotype is independent of the plastome. The experiments provide additional information on competition between megaspores and embryo sacs in the ovule.     

Early signs of disruption of wheat anther development associated with the induction of male sterility by meiotic-stage water deficit

 Water deficit during meiosis in microspore mother cells of wheat (Triticum aestivum L.) induces male sterility, which reduces grain yield. In plants stressed during meiosis and then re-watered, division of microspore mother cells seems to proceed normally, but subsequent pollen development is arrested. Stress-affected anthers generally lack starch. We employed light microscopy in conjunction with histochemistry to compare the developmental anatomy of water-stress-affected and normal anthers. The earliest effects of stress, detectable between meiosis and young microspore stages, were the degeneration of meiocytes, loss of orientation of the reproductive cells, and abnormal vacuolization of tapetal cells. Other effects observed during subsequent developmental stages were deposition of starch in the connective tissue where it is normally not present, hypertrophy of the middle layer or endothecial cells, and deposition of sporopollenin-like substances in the anther loculus. The resulting pollen grains lacked both starch and intine. These results suggest that abnormal degeneration of the tapetum in water-stressed anthers coupled with a loss of orientation of the reproductive cells could be part of early events leading to abortion of microspores. Received: 19 July 1996 / Revision accepted: 6 November 1996     

Mitochondrial genome diversity in connection with male sterility in Allium schoenoprasum L.

Mitochondrial genome diversity in chives (Allium schoenoprasum L.) was investigated with respect to different forms of male sterility. Cytoplasmic male-sterile (CMS) and restored genotypes of the known CMS system, compared to plants of the wi-, the st1- and the st2-sterility types and additional fertile plants of different origin were examined by means of RFLP analyses using mitochondrial gene probes. Besides the (S)-cytoplasm of the CMS system four additional cytoplasms were distinguished that differed in the organisation of their mitochondrial genomes. There is consequently a high degree of variability of the mitochondrial genome in chives, especially when compared with the closely related onion. A possible function of the atp9 gene in generating the different cytoplasm types of chives is discussed in relation to the origin of known CMS sequences in other plant species. The existence of different cytoplasm types offers the opportunity for further characterisation of the wi-, st1- and st2-sterility systems with respect to cytoplasmic factors which might be causally related to them. Whether these new sterilities are CMS or GMS (genic male sterilities) is of interest to plant breeders in order that restrictions on the genetic basis used in hybrid seed production be avoided. Received: 6 July 1999 / Accepted: 6 September 1999     

植物细胞核雄性不育相关bHLH转录因子研究进展

雄性不育广泛存在于种子植物中。植物雄性不育不仅是植物生殖发育研究的重要内容,同时也可作为杂种优势利用的有效工具,因而具有重要的理论和应用价值。bHLH转录因子家族是植物中成员最多的转录因子家族,在植株的整个生长发育过程中起着重要的调控作用。本文介绍了拟南芥、水稻、玉米等几种重要模式植物bHLH转录因子调控雄蕊发育的作用机制,并重点阐述其功能异常引起细胞核雄性不育的分子机制,以期为作物育种与理论研究提供参考。