单显性细胞核雄性不育油菜花雷表达蛋白比较研究

以受1对显性基因控制的单显性细胞核雄性不育油菜为材料,运用蛋白双向电泳技术对初花期不育花蕾和可育花蕾中蛋白质表达差异进行分析.结果表明,可育花蕾中表达的蛋白质总点数高于不育系.不育花蕾和可育花蕾中共有的蛋白点为223个,不育花蕾特有的蛋白质点数为103个,而可育花蕾中特有的蛋白质点数为160个.可育花蕾中表达的特有蛋白质分子量主要分布在60kD以下的小分子量区域,30kD尤为丰富;不育花蕾中表达的特有蛋白按分子量分布相对均匀.两者表达特有蛋白都相对集中在pI6.0～7.5区域,多为中性蛋白.     

单显性细胞核雄性不育油菜花蕾表达蛋白比较研究

以受1对显性基因控制的单显性细胞核雄性不育油菜为材料,运用蛋白双向电泳技术对初花期不育花蕾和可育花蕾中蛋白质表达差异进行分析.结果表明,可育花蕾中表达的蛋白质总点数高于不育系.不育花蕾和可育花蕾中共有的蛋白点为223个,不育花蕾特有的蛋白质点数为103个,而可育花蕾中特有的蛋白质点数为160个.可育花蕾中表达的特有蛋白质分子量主要分布在60kD以下的小分子量区域,30kD尤为丰富;不育花蕾中表达的特有蛋白按分子量分布相对均匀.两者表达特有蛋白都相对集中在pI6.0～7.5区域,多为中性蛋白.     

紫稻细胞质雄性不育系叶片全蛋白双向电泳分析

通过对几种不育系叶片全蛋白双向电泳图谱分析证明;紫稻不育系具有不同于野败型,红莲型和马协型不育系的蛋白图谱,说明其可能是一种新的细胞质质源,同时,紫稻不育系与保持系蛋白图谱之间在三叶期和分蘖期时差异均不明显,但各材料本身蛋白图谱在两个不同时期之间差异很大,不育系与保持系图谱表现出的蛋白质（多肽）的差异,可能与不育系雄性不育有关。     

光周期诱导的光敏核不育水稻农垦58S叶蛋白变化及其效应研究

本文用同位素标记和双向电泳方法,分析了光敏核不育水稻农垦58S敏感期的叶蛋白变化,共得到200个左右的蛋白质（肽）点,分布在P14.1-7.2,MW 10--95KD之间。其中．有4个蛋白质点（点1-4）的浓度明显受光周期影响而发生量的变化,而点5-8则是在长日照诱导下出现的新蛋白点。另外,在短日条件下稳定存在的点9,在长日照下却完全检测不到。光敏核不育水稻的育性转换,可能与这些蛋白质的变化有关。     

水稻光(温)敏核不育系与核质互作不育系的遗传关系剖析

1991─1992年,分析了7个光(温)敏核不育系与15 个核质互作不育系杂交F1及部分F2植株在长日照和短日照条件下的育性表现,结果清楚地表明,有些光敏核不育系能够保持核质互作不育系的雄性不育性,有些光(温)敏核不育系则能够恢复或部分恢复; 有些光敏核不育系对某一核质互作不育系具有保持能力,对另一核质互作不育系则具有恢复能力; 并初步推测光(温)敏核不育基因与核质互作不育基因是独立发生的,当核质互作不育系中细胞质和细胞核的隐性不育基因一起作用时,能够掩盖光(温)敏不育基因及其育性恢复基因的表达。     

溴氰菊酯胁迫下小菜蛾幼虫体内差异表达蛋白的分析

小菜蛾Plutella xylostella L.是世界性十字花科蔬菜的主要害虫, 已对多种杀虫剂产生抗性, 其中以对拟除虫菊酯类杀虫剂的抗性发展最快。溴氰菊酯是拟除虫菊酯杀虫剂中杀虫毒力最强的品种。我们前期的研究发现, 小菜蛾溴氰菊酯敏感品系（DS）和抗性品系（DR）成虫期的蛋白质双向电泳(2-DE)图谱存在显著差异。本研究通过双向电泳技术从小菜蛾4龄幼虫中分离出89个有明显差异的蛋白点, 从中选出30个进行串联质谱(MALDI-TOF-MS)实验, 并利用蛋白质数据库检索这些在抗性品系中表达而在敏感品系中不表达或者不同品系中差异表达的蛋白质的归属、 性质和功能, 最终成功鉴定出10个蛋白。对其中的3个基因进行了荧光定量PCR验证, 发现这些蛋白质在mRNA水平的表达与在蛋白水平的表达是一致的。这些在溴氰菊酯胁迫下差异表达的蛋白为研究溴氰菊酯的作用靶标和作用机理, 以及筛选与其抗性相关的蛋白质提供了依据。     

普通小麦D2型CMS系的育性基因及其遗传特性

通过研究普通小麦D^2型CMS－育性恢复体系中育性基因的种类及其遗传特性。结果表明：（1）D^2型不育系具有较好的不育性保持与恢复特征,在一般的普通小麦品种（系）中具有广泛的恢复（基因）源、可恢复度高（恢复度超过50％的品种或品质占到33．61％）,也能较容易地转育出新的不育系（完全保持不育性的品种或品系占到25．21％）,这一特征明显优于现有T、K、V型等不育系。（2）D^2型不育系的不育性受核内不育基因和抑制基因控制,相应的核基因型分为Al（不育基因）、A2（不育基因＋抑制基因）两类;恢复纱的恢复性受核内主效恢复基因、微效恢复基因和抑制基因控制,相应的核基因型分为C1（主效恢复基因）、C2(驻效恢复基因＋微效恢复基因）、C3（微效恢复基因）、C4（主效恢复基因＋抑制基因）、C5（主效恢复基因＋微效恢复基因＋抑制基因）、C6（微效恢复基因＋抑制基因）6种。环境条件的变化对育性基因、尤其是微效恢复基因和抑制基因的遗传效应有不同程度的影响。D^2型不育有效杂交组合的模型为：A1＋C1`A1 C2、A2＋C2。（3）D^2型不育系等位恢复基因的遗传表现为不完全显性,非等位恢复基因的遗传表现出积效应,这正是强恢复系德育的理论依据之一。     

光周期诱导的光敏核不育水稻农垦58S叶蛋白变化及其效应研究

本文用同位素标记和双向电泳方法,分析了光敏核不育水稻农垦58S敏感期的叶蛋白变化,共得到200个左右的蛋白质(肽)点,分布在PI4.1—7.2,MW10—95KD之间。其中,有4个蛋白质点(点1—4)的浓度明显受光周期影响而发生量的变化,而点5—8则是在长日照诱导下出现的新蛋白点。另外,在短日条件下稳定存在的点9,在长日照下却完全检测不到。光敏核不育水稻的育性转换,可能与这些蛋白质的变化有关。     

YS型小麦温敏雄性不育系A3017育性相关基因的SSH分析

以人工气候箱控温条件下YS型小麦温敏不育系A3017的不育幼穗和可育幼穗为材料,分别构建了不育和可育条件下基因特异表达的抑制消减杂交cDNA文库.在两个库中分别随机挑选100个阳性克隆进行测序,经BLAST序列比对分析,共获功能已知的EST 78条.比较发现,不育条件下与细胞质相关的基因表达数量(63.4%)远高于可育条件下的基因数量(35.1%),参与蛋白质合成、蛋白质修饰/加工/储藏、转运和信号传递过程基因的比例也均高于可育条件下的,而参与能量代谢过程基因的比例则明显偏低.分析认为,在不育和可育条件下的基因表达差异可能是导致温敏雄性不育系育性变化的关键,这为进一步从分子基础上揭示YS型小麦温敏雄性不育系育性转换机制奠定了基础.     

高粱热激蛋白(HSPs)的电泳分析与雄性不育性

本文以高粱雄性不育系为材料进行了热激蛋白的研究。苗期单向电泳表明,保持系在热激(40℃)过程中可溶性蛋白有24条带,对照(28℃)22条,出现2条带差异,不育系在热激后有33条带,比对照28条多5条,且有4条加强带,共9条带产生差异。苗期双向电泳,不育系在热激后比对照有19点产生差异,保持系热激后与对照比较有6点产生差异。表明双向电泳揭示了热激后蛋白质在分子量和电性方面有差异。花粉母细胞期幼穗的热激蛋白,不育系对照可溶性蛋白仅有3条带,热激后有11条,8条带有差异。保持系对照有10条带,热激后有15条,1条加强,共6条带有差异。幼穗期不育系蛋白质突出的缺少19kd以上的大分子量的带。从个体发育看,不育系由苗期可溶性蛋白比保持系带数多,发育到花粉母细胞期比保持系突出地减少,尤其是大分子量的蛋白带消失,热激后明显地增加了与可育的保持系相同的蛋白成份,表明不育系有其特殊的基因调控。     

Mitochondrial aldehyde dehydrogenase activity is required for male fertility in maize

Some plant cytoplasms express novel mitochondrial genes that cause male sterility. Nuclear genes that disrupt the accumulation of the corresponding mitochondrial gene products can restore fertility to such plants. The Texas (T) cytoplasm mitochondrial genome of maize expresses a novel protein, URF13, which is necessary for T cytoplasm-induced male sterility. Working in concert, functional alleles of two nuclear genes, rf1 and rf2, can restore fertility to T cytoplasm plants. Rf1 alleles, but not Rf2 alleles, reduce the accumulation of URF13. Hence, Rf2 differs from typical nuclear restorers in that it does not alter the accumulation of the mitochondrial protein necessary for T cytoplasm-induced male sterility. This study established that the rf2 gene encodes a soluble protein that accumulates in the mitochondrial matrix. Three independent lines of evidence establish that the RF2 protein is an aldehyde dehydrogenase (ALDH). The finding that T cytoplasm plants that are homozygous for the rf2-R213 allele are male sterile but accumulate normal amounts of RF2 protein that lacks normal mitochondrial (mt) ALDH activity provides strong evidence that rf2-encoded mtALDH activity is required to restore male fertility to T cytoplasm maize. Detailed genetic analyses have established that the rf2 gene also is required for anther development in normal cytoplasm maize. Hence, it appears that the rf2 gene was recruited recently to function as a nuclear restorer. ALDHs typically have very broad substrate specificities. Indeed, the RF2 protein is capable of oxidizing at least three aldehydes. Hence, the specific metabolic pathway(s) within which the rf2-encoded mtALDH acts remains to be discovered.     

Genetic architecture of male fertility restoration of <Emphasis Type="Italic">Triticum timopheevii</Emphasis> cytoplasm and fine-mapping of the major restorer locus <Emphasis Type="Italic">Rf3</Emphasis> on chromosome 1B

Key message

Restoration of fertility in the cytoplasmic male sterility-inducing Triticum timopheevii cytoplasm can be achieved with the major restorer locus Rf3 located on chromosome 1B, but is also dependent on modifier loci.

Abstract

Hybrid breeding relies on a hybrid mechanism enabling a cost-efficient hybrid seed production. In wheat and triticale, cytoplasmic male sterility based on the T. timopheevii cytoplasm is commonly used, and the aim of this study was to dissect the genetic architecture underlying fertility restoration. Our study was based on two segregating F₂ triticale populations with 313 and 188 individuals that share a common female parent and have two different lines with high fertility restoration ability as male parents. The plants were cloned to enable replicated assessments of their phenotype and fertility restoration was evaluated based on seed set or staining for pollen fertility. The traits showed high heritabilities but their distributions differed between the two populations. In one population, a quarter of the lines were sterile, conforming to a 3:1 segregation ratio. QTL mapping identified two and three QTL in these populations, with the major QTL being detected on chromosome 1B. This QTL was collinear in both populations and likely corresponds to Rf3. We found that Rf3 explained approximately 30 and 50% of the genotypic variance, has a dominant mode of inheritance, and that the female parent lacks this locus, probably due to a 1B.1R translocation. Taken together, Rf3 is a major restorer locus that enables fertility restoration of the T. timopheevii cytoplasm, but additional modifier loci are needed for full restoration of male fertility. Consequently, Rf3 holds great potential for hybrid wheat and triticale breeding, but other loci must also be considered, either through marker-assisted or phenotypic selection.

     

二角型非1B/1R 小麦CMS不育恢复体系的建立

通过对具有二角山羊草细胞质的1B/1R型小麦雄性不育系m s(A e.bicorn is)-5-1回交置换,获得了新型二角山羊草细胞质小麦雄性不育系m s(A e.bicor)-V 9125和m s(A e.bicor)-M 853.利用染色体制片、分子标记、原位杂交和酸性聚丙烯酰胺凝胶电泳对其保持系V 9125和M 853进行分子细胞遗传学检测,并对其育性恢复性机理进行了初步研究.结果表明:(1)V 9125和M 853均为非1B/1R易位系.V 9125为小麦-簇毛麦易位系,m s(A e.bicor)-V 9125是二角山羊草细胞质与普通小麦核背景中簇毛麦外源染色体互作产生的一类新的核质互作不育系.(2)M 853为小麦-滨麦易位系,m s(A e.bicor)-M 853是二角山羊草细胞质与普通小麦核背景中滨麦外源染色体互作产生的一类新的核质互作不育系.(3)利用一些普通小麦品种(系)与这两个不育系杂交,m s(A e.bicor)-V 9125和m s(A e.bicor)-M 853仅与T 6-3杂交育性得到恢复,且恢复度较高,变异小,而与其它大部分普通小麦杂交F1表现雄性不育,且扬花期花药不外露.而m s(A e.bicor)-5-1与包括T 6-3在内的普通小麦品种(系)杂交F1育性得到不同程度的恢复.从而得出结论,二角山羊草细胞质与小麦细胞核的互作存在两个不同的不育-恢复系统.     

Comparative analysis of fertility restoration genes for WA,Y, and DA cytoplasmic male sterility in rice

Rice chromosome single segment substitution line (SSSL) W23-19-06-06-11 with the genotype Rf3Rf3/Rf4Rf4, a strong restorer line for wild-abortive (WA) cytoplasmic male sterility (CMS), was recently identified from the SSSL library. To investigate the genetic mode of Rf genes and the genetic relationship among WA, yegong (Y), and dwarf-wildabortive (DA) CMS systems, the plants derived from three BC3F2 populations involving W23-19-06-06-11 and the three CMS lines, that carried the Rf3Rf3/Rf4Rf4, Rf3Rf3/rf4rf4, and rf3rf3/Rf4Rf4 genotypes and WA-, Y-, and DA-CMS cytoplasm, were selected and their pollen and spikelet fertility were evaluated. The results show that the genetic effect displayed a trend of Y-CMS > WA-CMS > DA-CMS in the genetic background of W23-19-06-06-11, the effect of Rf4 appeared to be slightly larger than that of Rf3, and their effects were additive for the three CMS systems. Two pairs of dominant genes governed the fertility restoration in pollen and spikelet in the W23-19-06-06-11 which indicates that the genetic mode of the Rf genes was a qualitative character for the three CMS systems.     

Recovery of Heritable, Transposon-Induced, Mutant Alleles of the Rf2 Nuclear Restorer of T-Cytoplasm Maize

T (Texas) cytoplasm is associated with a mitochondrial disruption that is phenotypically expressed during microsporogenesis resulting in male sterility. Restoration of pollen fertility in T-cytoplasm maize is controlled by dominant alleles at two unlinked, complementary, nuclear-encoded genes, rf1 and rf2. As a first step in the molecular isolation of the rf2 gene, 178,300 gametes derived from plants that carried the Mutator, Cy or Spm transposon families were screened for rf2 mutant alleles (rf2-m) via their inability to restore pollen fertility to T-cytoplasm male-sterile maize. Seven heritable rf2-m alleles were recovered from these transposon populations. Pedigrees and restriction fragment length polymorphism (RFLP)-based analyses indicated that all seven rf2-m alleles were derived independently. The ability to obtain rf2-m derivatives from Rf2 suggests that Rf2 alleles produce a functional product necessary to restore pollen fertility to cmsT. Molecular markers flanking the rf1 and rf2 loci were used to decipher segregation patterns in progenies segregating for the rf2-m alleles. These analyses provided preliminary evidence of a weak, third restorer gene of cmsT that can substitute for Rf1.     

RFLP mapping of the maize gametophytic restorer-of-fertility locus (rf3) and aberrant pollen transmission of the nonrestoring rf3 allele

 Cytoplasmic male sterility (CMS) is the maternally inherited inability to produce functional pollen. The Rf3 allele of the nuclear gene rf3 gametophytically restores male fertility to maize plants with the S-type of CMS. The rf3 locus is on the long arm of maize chromosome two (2L). Using 2L RFLPs and three-point mapping analysis we showed that the rf3 locus is located an estimated 4.3 cM distal to the whp locus and 6.4 cM proximal to the bnl17.14 locus. This information was used in combination with RFLPs on two additional maize chromosomes to show that Rf3/rf3 CMS-S plants may aberrantly transmit the nonrestoring allele, rf3, through the male gametophyte. Received: 30 September 1996/Accepted: 21 March 1997     

Intrachromosomal mapping of the nucleolar organiser region relative to three marker loci on chromosome 1B of wheat (Triticum aestivum)

Summary Restriction enzyme digestion of the ribosomal RNA genes of the nucleolar organisers of wheat has revealed fragment length polymorphisms for the nucleolar organiser on chromosome 1B and the nucleolar organiser on 6B. Variation between genotypes for these regions has also been demonstrated. This variation has been exploited to determine the recombination frequency between the physically defined nucleolar organiser on 1B (designatedNor1) and other markers; two loci,Glu-B1 andGli-B1 which code for endosperm storage proteins andRf3, a locus restoring fertility to male sterility conditioned byT. timopheevi cytoplasm.Gli-B1 andRf3 were located on the short-arm satellite but recombine with the nucleolar organiser giving a gene order ofNor1 — Rf3 — Gli-B1. Glu-B1 is located on the long arm of 1B but shows relatively little recombination withNor1, which is, in physical distance, distal on the short arm. This illustrates the discrepancy between map distance and physical distance on wheat chromosomes due to the distal localisation of chiasmata. The recombination betweenNor1 andRf3 indicates that, contrary to previous suggestions, fertility restoration is not a property of the nucleolar organiser but of a separate locus.