植物雄性不育基因的研究进展

本文概述了植物雄性核不育基因的分子标记及其定位,综述了植物细胞质雄性不育中不育系与保持系在叶绿体和线粒体基因组的结构、转录和翻译产物方面的差异以及和雄性不育之间的可能关系,以及恢复系中的恢复基因分子水平的研究现状;讨论了环境条件光周期和温度对雄性不育的影响在分子水平上的研究现状,指出了植物雄性不育基因研究方面存在的问题和解决贩思路。     

雄性不育在作物杂种优势中的应用途径分析

雄性不育是植物雄性细胞或生殖器官丧失生理机能的现象,该现象的利用大大提高了杂交种生产的效率。植物雄性不育包含细胞质雄性不育、不受环境影响的核雄性不育、光温敏型雄性不育及化学诱导的雄性不育。这些不育类型也已经被以三系或二系的方式应用于很多作物的杂交种生产中。综述了雄性不育各个途径的研究进展及其在作物杂种优势中的应用。     

向日葵CMS育性恢复的研究

向日葵细胞质雄性不育（Ｃｙｔｏｐｌａｓｍｉｃｍａｌｅｓｔｅｒｉｌｉｔｙ,ＣＭＳ）育性恢复的机理是非常复杂的。运用遗传学和分子生物学方法,分析了具代表性的４种不同细胞质类型的ＣＭＳ育性被恢复的频率和２０种向日葵自交系对１９种ＣＭＳ的恢复能力及个别ＣＭＳ植株自发恢复的原因。实验结果表明,４种ＣＭＳ品系育性被恢复的频率分别为５８．８％．５６．３％．１１．８％和０％．２０种自交系的恢复力为５．９－７５．０％。部分ＣＭＳ品系和大多数自交系含有恢复基因,恢复基因的数量及类型决定了ＣＭＳ品系被恢复的程度及自交系的恢复能力。同时,提出并证实了线粒体不育基因变异是导致ＡＲＧ１ＣＭＳ植株自发恢复育性的主要原因。     

Molecular characterization of a gene encoding a cysteine-rich protein preferentially expressed in anthers of Lycopersicon esculentum

The tomato (Lycopersicon esculentum Mill.) cDNA clone TomA5B was isolated by differential screening of a cDNA library prepared from anthers at late meiosis to tetrad formation. The 5B gene is present in a single copy in the tomato genome. Expression is developmentally regulated and tissue specific. RNA accumulation was detected from premeiosis through tetrad release in the tapetal cell layer of the anther with low levels of RNA detected in petals and early stages of pistil development. The protein deduced from the DNA sequence analysis is predicted to have a molecular mass of 11.1 kDa and a secretory signal sequence, suggesting it is a secreted protein. The deduced 5B protein has a pattern of cysteine residues that is similar to other proteins that have stamen-specific expression and to a superfamily of seed proteins. The 5B protein is unique in that there is no amino acid sequence similarity to other proteins beyond the similar cysteine motif.     

Premature Tapetum Degeneration: a Major Cause of Abortive Pollen Development in Photoperiod Sensitive Genic Male Sterility in Rice

Photoperiod-sensitive genic male-sterile (PSGMS) rice ( Oryza sativa L.), a natural mutant found in the rice cultivar Nongken 58, is very useful for the development of hybrid rice cultivars. Despite its widespread use in breeding programs, the initial stage of the abortive development of PSGMS rice and the possible cytological mechanisms of pollen abortion have not been determined. In the present study, a systematic cytological comparison of the anther development of PSGMS rice with its normal fertile counterpart is conducted. The results show that pollen abortion in PSGMS rice first occurs before the pollen mother cell (PMC) stage, and continues during the entire process of pollen development until pollen degradation. The abortive process was closely associated with the abnormal behavior of the tapetum. Although tapetum degeneration in PSGMS rice initiates already at the PMC stage, it proceeds slowly and does not complete until the breakdown of the pollen. Such cytological observations were supported by the results of the TUNEL (TdT-mediated dUTP Nick End Labeling) assay, which detects DNA fragmentation resulting from programmed cell death (PCD), indicating that the premature tapetum degeneration is in the process of PCD.     

Characterization and Use of Male Sterility in Hybrid Rice Breeding

The hybrid rice （Oryza sativa L.） breeding that was Initiated In China in the 1970s led to a great improvement in rice productivity. In general, It increases the grain yield by over 20% to the inbred rice varieties, and now hybrid rice has been widely introduced into Africa, Southern Asia and America. These hybrid varieties are generated through either three-line hybrid and two-line hybrid systems; the former is derived from cytoplasmic male sterility （CMS） and the latter derived from genlc male sterility （GMS）. There are three major types of CMS （HL, BT and WA） and two types of GMS （photoperlod-sensitlve （PGMS） and temperature-sensitive （TGMS））. The BT- and HL-type CMS genes are characterized as orf79 and orfH79, which are chimeric toxic genes derived from mltochondrial rearrangement. Rf3 for CMS-WA Is located on chromosome 1, while Rf1, Rf4, Rf5 and Rf6 correspond to CMS-BT, CMSoWA and CMS- HL, located on chromosome 10. The Rfl gene for BT-CMS has been cloned recently, and encodes a mltochondriatargeted PPR protein. PGMS Is thought to be controlled by two recessive loci on chromosomes 7 and 12, whereas nine recessive alleles have been identified for TGMS and mapped on different chromosomes. Attention Is still urgently needed to resolve the molecular complexity of male sterility to assist rice breeding.     

玉米细胞质雄性不育及其育性恢复基因的研究进展

玉米是杂种优势利用最成功的作物之一,采用细胞质雄性不育(CMS)进行玉米杂交种生产已成为杂种优势利用的有力工具。CMS是由于细胞质和细胞核的基因表达产物的不协调而产生的不育性,可被核基因组中的恢复基因恢复。根据育性恢复专效性,玉米CMS材料主要分为T、C和S三种类型。综述了这三种类型不育及其恢复基因的研究进展,分析了在不育化制种中的应用情况。     

植物细胞质雄性不育及其育性恢复的分子基础

植物细胞质雄性不育是广泛存在于高等植物中的现象,其表现为母性遗传、花粉败育,但雌蕊正常。细胞质雄性不育在杂交种子生产中起着重要作用,研究其分子作用机制有利于更有效地利用细胞质雄性不育。随着一些不育基因和恢复基因相继被克隆,人们对一些细胞质雄性不育和恢复系统的分子作用机理已经有一定了解。本文综述了近年来对植物细胞质雄性不育基因和恢复基因作用机理研究的进展。     

一个新的小麦核不育材料的发现和鉴定

从轮选可育株衍生的后代中发现一个雄性不育株,即P740不育材料。在P7 40不育材料第1、2、3次姊妹交后代中,可育株与不育株的分离比分别符合4:1、 7:5和15:13。在第2次姊妹交后代中可育株产生的40个株系中,7个株系的育性分离比是15:1,33个株系是3:1,二者约分别占1/7和6/7。以上结果表明,P740是一个双隐性基因控制的核雄性不育材料。本文提出了计算隐性核不育材料姊妹交后代育性分离比率的公式。 Abstract:A male sterile material was found in the progeny derived from a fertile plant in the population of recurrent selection.The ratios of fertile to sterile plants in three sib-mating of the male sterile material with fertile plants were to 4:1,7:5 and 15:13,respcctively.In the 40 plant lines derived from the fertile plants of second sib-mating population,33 lines approximated to the segregation ratio of 3(fertile):a double recessive nuclear male sterile material.A genetic pattern for calculation of fertile segregation in sib-mating progenies of recessive nuclear male sterile material was advanced in this paper.     

水稻突变体与功能基因组学

突变体是功能基因组学研究的重要材料。本文综述了水稻突变体的创制方法、变异的机制、水稻突变体,基因分类和数量、克隆的水稻重要基因的研究进展,包括1698个水稻突变体,基因的分类和43个克隆的水稻突变体基因,并提出了利用水稻突变体的展望。     

雄性不育相关基因msLTP的反义RNA验证

根据普通白菜雄性不育相关的脂质转移蛋白基因(msLTP)的cDNA序列设计引物,从普通白菜花蕾的cDNA中扩增出312bp的片段,然后将该片段连接至双元载体pBI12l中,得到反义RNA植物表达载体并导入农杆菌LBA4404菌株中,通过农杆菌介导法转化菜心;利用PCR和Southern blot分析检测得到了25株转基因植株,转基因植株的花粉部分畸形或空瘪,花粉离体萌发率为38.56%,较未转化植株的萌发率(76.32%)降低了37.76个百分点.研究表明,反义RNA技术使msLTP基因沉默而导致了菜心转基因植株的部分花粉发育不良,说明msLTP基因在普通白菜和菜心等花粉发育中具有重要作用.     

小麦光温敏雄性不育相关基因的DDRT－PCR分析及功能预测

 以小麦光温敏雄性不育系BS210为试材,分别在北京顺义（可育环境）和安徽阜阳（不育环境）两种诱导育性表现的生态环境下种植,并于雌雄蕊分化期、药隔形成期、花粉母细胞形成期、四分体期和单核期5个时期提取小穗组织的RNA,利用DDRT-PCR方法分离不育系在光、温因子诱导下的差异表达mRNA,并通过反向Northern 进行验证.对获得的20条差异表达的EST进行了测序和BLAST分析,得到了4个候选基因的片段,对其进行5′Race 扩增、序列分析及功能预测.结果表明：它们分别与水稻的DNA修复重组蛋白基因rad50、小麦穗部表达的LRR重复序列型类受体激酶基因、玉米叶片坏死斑点L1s1基因的序列相似性分别为89%、89%和88%,另外还筛选到1个未知功能的新基因片段,它和rad50分别在可育和不育环境下表达的mRNA具有相同的5′端编码区,但3′端非编码区     

甘蓝型油菜新型细胞质雄性不育类型NRO4270A的鉴定

甘蓝型油菜细胞质雄性不育系(CMS)NRO4270A是从萝卜甘蓝(Raphanus brassica,RRCC)与甘蓝型油菜(Brassica napus,AACC)远缘杂交后代中发现的一种CMS类型,该研究通过育性调查、恢保关系测定、花药发育的显微观察和线粒体DNA的RFLP分析对NRO4270A不育系进行鉴定。结果表明:(1)通过连续5年在冬油菜区和春油菜区的育性调查,发现NRO4270A不能产生花粉,败育彻底,不育性稳定,不受温度、光照等环境条件的影响,不育率和不育度均为100%。(2)将NRO4270A、pol、ogu、kos、hau等CMS分别与pol、ogu恢复系以及‘华双4号’测交,显示NRO4270A与pol、ogu、kos等不育系统的恢保关系明显不同。(3)通过对NRO4270A与保持系NRO4270B花药发育的显微结构观察和比较,表明NRO4270A花粉的败育时期为四分体期至单核花粉晚期,败育特点为:单核小孢子时期,单核花粉粒不能形成外壁,花药绒毡层液泡化,并逐渐膨大增厚,最后绒毡层和小孢子彻底消失,花粉囊变为空腔,花药完全败育。(4)利用18个探针/酶组合(由6个线粒体基因atp1、atp6、atp9、orf222、cox2、cob探针和3种限制性内切酶EcoRⅠ、BamHⅠ和HindⅢ组成)对NRO4270A、pol、ogu、kos、hau等CMS以及NRO4270B和‘华双5号’的线粒体DNA进行RFLP分析,在11个探针/酶组合中,NRO4270A均显示与其他材料明显不同的带型,表明其为新型的不育胞质类型。NRO4270A的发现和利用将有利于克服中国油菜主要CMS类型对环境的敏感性和生产上不育胞质类型单一化等问题。     

植物细胞质雄性不育

近年来国内外对植物细胞质雄性不育（cytoplasmic male sterility,CMS）在细胞学、生理生化及分子遗传学等方面的研究又取得了新的进展。特别是对育性恢复基因的定位、克隆的研究已取得一定突破,发现编码含保守PPR（pentatricopeptide repeat）模体蛋白的基因在多种植物中与育性恢复密切相关。     

水稻广亲和性和胞质雄性不育恢复性的遗传分析

对经花培育成的广亲和恢复系ＴＧ７、ＴＧ８的遗传分析表明：ＴＧ７、ＴＧ８均带有１对广亲和基因,与ＣＰＳＬＯ１７、０２４２８的广亲和基因等位。广亲和基因与雄性不育恢复基因表现为独立遗传,野败型（籼）和滇－Ｉ型（粳）不育胞质的育性恢复基因非等位。ＴＧ７和ＴＧ８分别带有２对野败不育胞质和１对滇－Ｉ型不育胞质的恢复基因,分别来自亲本明恢６３和ＣＰＳＬＯ１７。     

作物雄性不育性在育种中的应用概评

概述总结了作物雄性不育性的类别与遗传特点。雄性不育性的遗传机理涉及细胞质遗传的现象,目前已初步探明玉米C群不育系的胞质基因可能是atp6-c,芝麻不育胞质基因拟为atpA。雄性不育化杂交种在实践中主要应用于玉米、水稻和蔬菜中。尽管现有近交理论、DNA甲基化效用、水稻胞质与核不育系遗传等理论提出,雄性不育化育种的基本理论尚需进一步探讨。在雄性不育化育种技术上,要逐步解决难点作物,如小麦、荞麦、菜豆等的不育化育种问题。     

Fertile Arabidopsis cyp704b1 mutant,defective in sporopollenin biosynthesis,has a normal pollen coat and lipidic organelles in the tapetum

The exine acts as a protectant of the pollen from environmental stresses, and the pollen coat plays an important role in the attachment and recognition of the pollen to the stigma. The pollen coat is made of lipidic organelles in the tapetum. The pollen coat is necessary for fertility, as pollen coat-less mutants, such as those deficient in sterol biosynthesis, show severe male sterility. In contrast, the exine is made of sporopollenin precursors that are biosynthesized in the tapetum. Some mutants involved in sporopollenin biosynthesis lose the exine but show the fertile phenotype. One of these mutants, cyp704b1, was reported to lose not only the exine but also the pollen coat. To identify the cause of the fertile phenotype of the cyp704b1 mutant, the detailed structures of the tapetum tissue and pollen surface in the mutant were analyzed. As a result, the cyp704b1 mutant completely lost the normal exine but had high-electron-density granules localized where the exine should be present. Furthermore, normal lipidic organelles in the tapetum and pollen coat embedded between high-electron-density granules on the pollen surface were observed, unlike in a previous report, and the pollen coat was attached to the stigma. Therefore, the pollen coat is necessary for fertility, and the structure that functions like the exine, such as high-electron-density granules, on the pollen surface may play important roles in retaining the pollen coat in the cyp704b1 mutant.     

Rapid Macrosatellite Evolution Promotes X-Linked Hybrid Male Sterility in a Feline Interspecies Cross

The sterility or inviability of hybrid offspring produced from an interspecific mating result from incompatibilities between parental genotypes that are thought to result from divergence of loci involved in epistatic interactions. However, attributes contributing to the rapid evolution of these regions also complicates their assembly, thus discovery of candidate hybrid sterility loci is difficult and has been restricted to a small number of model systems. Here we reported rapid interspecific divergence at the DXZ4 macrosatellite locus in an interspecific cross between two closely related mammalian species: the domestic cat (Felis silvestris catus) and the Jungle cat (Felis chaus). DXZ4 is an interesting candidate due to its structural complexity, copy number variability, and described role in the critical yet complex biological process of X-chromosome inactivation. However, the full structure of DXZ4 was absent or incomplete in nearly every available mammalian genome assembly given its repetitive complexity. We compared highly continuous genomes for three cat species, each containing a complete DXZ4 locus, and discovered that the felid DXZ4 locus differs substantially from the human ortholog, and that it varies in copy number between cat species. Additionally, we reported expression, methylation, and structural conformation profiles of DXZ4 and the X chromosome during stages of spermatogenesis that have been previously associated with hybrid male sterility. Collectively, these findings suggest a new role for DXZ4 in male meiosis and a mechanism for feline interspecific incompatibility through rapid satellite divergence.     

Modifying the pollen coat protein composition in Brassica

The interactions between pollen and stigma are essential for plant reproduction and are made possible by compounds, such as proteins and lipids, located on their surfaces. The pollen coat is formed in part by compounds synthesized in, and released from, the tapetum, which become transferred to the pollen coat late in pollen development. In the Brassicaceae the predominant proteins of the mature pollen coat are the tapetal oleosin-like proteins, which are highly expressed in, and ultimately transferred from, the tapetum. Here we report the modification of the protein composition of the pollen coat by the addition of an active enzyme which was synthesized in the tapetum. The marker enzyme beta-glucuronidase (GUS) was successfully targeted to the pollen coat in transgenic Brassica carinata plants expressing GUS translationally fused to a B. napus tapetal oleosin-like protein (BnOlnB;4). To our knowledge this is the first demonstration of the targeting of an enzyme to the pollen coat.     

Effects of pre-dispersal selection on offspring growth and survival in Erythronium grandiflorum

Effects of pre-seed-dispersal processes on offspring vigor were examined in Erythronium grandiflorum using manipulations of the number of pollen donors contributing to the pollen pool and comparisons of means and variances in offspring growth measurements. There were no effects of the number of donors on measures of pollen-tube growth, ovule abortion, seed set, mean seed weight, or seedling germination. Seeds from pollinations with only one donor produced corms that averaged 5% lighter after one season of growth and had lower overall survival after three years compared to corms from pollinations with either three or ten donors. Patterns of within- and among-family variance estimates for the different treatments were consistent with the hypothesis that less-vigorous offspring were eliminated prior to seed dispersal in the multiple-donor treatments. The difference in the growth of offspring from different treatments was apparently not due to pollen competition because pre-zygotic attrition of pollen tubes led to incomplete fertilization of ovules. Results from this study suggest that post-fertilization abortion of less-vigorous progeny, perhaps as a consequence of early-acting inbreeding depression, is responsible for the increase in the average vigor of offspring from multiply-sired fruits.