异源细胞质小麦耐盐性的研究

以粗厚山羊草（Ａｅｇｉｌｏｐｓ ｃｒａｓｓａ）细胞质小麦为材料,采用组织培养,水培、模拟盐池等方法,研究细胞质对小麦耐盐性的遗传效应,结果表明：粗厚山羊草细胞质可以不同程度地诱发小麦耐盐性产生变异。不同核质组合细胞质效应有一定差异,表现出特定的核质互作关系。部分异质系细胞水平与植株水平的耐盐性表现一致,特别是（Ａｅ．ｃｒａｓｓａ）－鉴２６的愈伤组织和幼苗的耐盐性表现均最突出。返青期和成熟期的鉴定结     

光周期敏感细胞质雄性不育小麦的研究

选用我们创建的Ｄ＾２型细胞质与普通小麦品种,经多年回交核置换获得的１５种异质系在,不同发育期置于武汉不同光长条件下,并春播于哈尔滨自然长日条件下,抽穗时套袋自产,进行碘染花分观察,成熟后考察自交结实率等。     

Structural and Expressional Variation Analyses of Mitochondrial Genomes Reveal Candidate Transcripts for the SV Cytoplasmic Male Sterility in Wheat (Triticum aestivum L.)

Common wheat (Triticum aestivum L.) is one of the most important crops,and intra-specific wheat hybrids have obvious heterosis in yield and protein quality.Therefore,utilization of hybrid wheat varieties offers an effective way to increase yield and nutrition.Cytoplasmic male sterility (CMS) systems are a useful genetic tool for hybrid crop breeding,and are ideal models for studying the genetic interaction and cooperative function of mitochondrial and nuclear genomes in plants (Schnable and Wise,1998;Hanson and Bentolila,2004).The breeding of hybrid wheat using male sterility caused by the cytoplasm of T.timopheevii has been studied since the early 1960's.But it is unsuccessful because there are some deficiencies in the practical application of this cytoplasm,including limited restoration resources,thin seeds,pre-harvest sprouting and lower germination rate (Wilson and Ross,1962).The Sv type of cytoplasmic male sterility (CMS-Sv) in wheat is general accessions for four types of CMS lines that were derived from four Aegilops species:Ae.kotschyi,Ae.variabilis,Ae.ventricosa,and Ae.bicornis.Based on the observation of alloplasmic lines produced in all possible combinations between 12 wheat nuclear genotypes and 47 cytoplasms of two related genera,Triticum (wheat) and Aegilops,Ogihara and Tsunewaki (1988) concluded that the D2-cytoplasm of Ae.crassa and its relatives,N-cytoplasm of Ae.uniaristata,and SV-cytoplasm of Ae.kotschyi and its relatives might be used as the alternative male sterile cytoplasm to replace the T.timopheevii cytoplasm for hybrid wheat breeding.Ikeguchi et al.(1999) proposed that spring-type hybrid wheat may be bred by combination of the 1BL-1RS chromosome and Ae.kotschyi cytoplasm with a new fertility-restorer gene discovered in a wheat variety Kitamiharu 48.Zhang et al.(1996) also proposed the use of CMS-Sv lines as the most effective male sterile cytoplasm.     

几类异质易恢复小麦雄性不育系杂种籽粒品质研究初报

利用具有粘果出羊草（Ae.kotschyi),易变山羊草（Ae.variabilis),偏凸山羊草（Ae.ventricosa)和二角山羊草（Ae.bicornis)异源细胞质小麦雄性不育系（以下简称粘,易,偏和二角型）组配成不同组合的杂种小麦,研究这4种不育系对杂种小麦籽粒品质的影响。结果表明,由它们所配制的杂种小麦在蛋白质含量,湿面筋含量,沉淀值等性状上均高于具有普通小麦细胞质的相同核型杂种小麦,尤其粘,易型某些组合在湿面筋含量,沉淀值两性状上差异达到显著水平。因此,利用这4种不育系很有希望培育出优势,高产的杂种小麦。     

Pistillody, homeotic transformation of stamens into pistil-like structures, caused by nuclear-cytoplasm interaction in wheat

Homeotic transformation of stamens into pistil-like structures (pistillody) has been observed in a cytoplasmic substitution (alloplasmic) line of wheat (Triticum aestivum L.) cv. Norin 26, which has the cytoplasm of a wild relative species, Aegilops crassa L. On the other hand, an alloplasmic line of wheat cv. Chinese Spring (CS) with Ae. crassa cytoplasm has normal flowers. This is due to the presence in the CS nucleus of a fertility-restoring gene, Rfd1. Deletion mapping analysis revealed that Rfd1 is located on the middle part of the long arm of chromosome 7B. To investigate the function of the Rfd1 gene by a loss-of-function strategy, we produced alloplasmic lines of CS ditelosomic 7BS [(cr)-CSdt7BS] and CS monotelodisomic 7BS [(cr)-CSmd7BS] with the Ae. crassa cytoplasm, and characterized their phenotypes. The line (cr)-CSdt7BS without Rfd1 exhibited pistillody in all florets, and also female sterility. Scanning electron microscopy of the young spikes revealed that the pistillody was induced at an early stage of stamen development. The pistillate stamens often developed incomplete ovule-like structures with integuments instead of tapetum and pollen grains. It is possible that MADS box genes are associated with the induction of pistillody, because the expression of wheat APETALA3 homologue (WAP3) was reduced in the young spikes of (cr)-CSdt7BS. In addition, a histological study indicated that the female sterility in (cr)-CSdt7BS is due to the abnormality of the ovule, which fails to form an inner epidermis and integuments in the chalaza region. The line (cr)-CSmd7BS, hemizygous for Rfd1, showed partial pistillody (51%) and restored female fertility up to 72%. These results suggest that the induction of both pistillody and ovule deficiency caused by the Ae. crassa cytoplasm is inhibited by the Rfd1 gene in a dose-dependent manner.     

粗厚山羊草（Ae.crassa）细胞质普通小麦核代换系在中国不同光温条件地点的雄性育性变异

5个普通小麦品系的粗厚山羊草4X,6X种细胞质的7个核代换系在中国昆明,元谋,杨陵,互助,依安5个不同纬度地点种植的自交结实率,结合各点光温条件的分析表明,粗厚山羊草细胞质普通小麦核代换系的雄性育性随日长增加而下降,6X种细胞质核代换系在日长≥14.28h开始出现完全雄性不育,4X种细胞质核代换系在日长≥15.38h开始出现完全雄性不育;该细胞质核代换系的雄性育性在日长因子主控下,对温度变化也表现一定敏感性。粗厚山羊草细胞质背景下不同核基因型对光,温条件的敏感性有显著差异。核质互作效应是这种敏感性的遗传方式,粗厚山羊草4X、6X种细胞质的光,温敏感性存在差异。6X种细胞质的光,温敏感性高于4X种细胞质。     

异细胞质小麦与黑麦、小黑麦杂交对性状和减数分裂行为的影响

本文利用17种异细胞质“中国春”小麦与黑麦,小黑麦杂交,回交,研究其性状与减数分裂行为的表现。首次观察到D2型Ae.crassa.4x细胞质对同源染色体配对有抑制作用,对部分同源染色体配对有促进作用,SV型Ae.kotschyi细胞质对同源染色体,部分同源配对均有抑制作用,S1型Ae.sharonesis细胞质对部分同源染色体配对有促进,还观察到G型细胞质T.timopheevi,T.zhukovskyi,D2型细胞质,Ae.crassa 可提高产生有功能雌配子数,首次合成G型,SV型,D2型细胞质雄性不育的八倍体小黑麦,D2型细胞质八倍体小黑麦是光敏性雄性不育,在15小时以上的长光照条件下表现不育,这对进一步了解异细胞质的作用,小黑麦杂种优势的利用和小黑麦的改良均有重要意义。     

Effects of alien cytoplasmic variation on carbon assimilation and productivity in wheat

In glasshouse studies of four alloplasmic wheat series, phenotypic characters were least affected when the recipient parent cytoplasm was replaced by donor cytoplasm of the S or D plasmatype. In the T. aestivum cv. 'Selkirk' series, cytoplasm substitution did not affect Pmax per unit leaf area, although the flag leaf area (and photosynthetic rate per leaf) of each alloplasmic line was greater than that of euplasmic 'Selkirk'. In field trials, all the D plasmatype alloplasmics tested produced more ears m^-2 than did euplasmic 'Selkirk'. The increased tiller number and leaf area of alloplasmic lines resulted in greater canopy light interception than euplasmic 'Selkirk' early in the season. This characteristic was associated with reduced weed populations under crops of alloplasmic 'Selkirk' lines grown under low-, but not high-input, agronomic regimes, with Ae. cylindrica- and Ae. ventricosa-'Selkirk' significantly outyielding alloplasmic 'Selkirk' under low-input conditions. The F2 populations from crosses between European wheat varieties and 'Selkirk' lines exhibited higher standard deviations for grain yield for alloplasmic than for euplasmic 'Selkirk', suggesting potential for selecting heterotic nuclear-cytoplasmic combinations with alien cytoplasms.     

Identification of a protein kinase gene associated with pistillody,homeotic transformation of stamens into pistil-like structures,in alloplasmic wheat

Homeotic transformation of stamens into pistil-like structures (called pistillody) has been reported in cytoplasmic substitution (alloplasmic) lines of bread wheat (Triticum aestivum) having the cytoplasm of a wild relative species, Aegilops crassa. Our previous studies indicated that pistillody is caused by alterations of the class B MADS-box gene expression pattern associated with mitochondrial gene(s) in the Ae. crassa cytoplasm. To elucidate the nuclear gene involved in the cross-talk between pistillody-related mitochondrial gene(s) and nuclear homeotic genes, we performed cDNA subtraction analysis using cDNAs derived from young spikes of a pistillody line and a normal line. As a result, we identified a protein kinase gene, WPPK1 (wheat pistillody-related protein kinase 1), which is upregulated in the young spikes of the pistillody line. RT-PCR analysis indicated that WPPK1 is strongly expressed in pistils and pistil-like stamens in the pistillody line, suggesting that it is involved in the formation of pistil-like stamens as well as pistils. The full-length cDNA sequence for WPPK1 showed high similarity with a flowering plant PVPK-1 protein kinase, and phylogenetic analysis indicated that it is a member of AGC group protein kinases. Furthermore, a phosphorylation assay indicated that it has protein kinase activity. In situ hybridization analysis revealed that WPPK1 is expressed in developing pistils and pistil-like stamens as well as in their primordia. These indicate that in the alloplasmic line, WPPK1 plays a role in formation and development of pistil-like stamens.     

QTL analysis of fertility-restoration against cytoplasmic male sterility in wheat

The cytoplasm of Triticum timopheevi causes cytoplasmic male sterility (CMS) in common wheat (T. aestivum) cv. 'Chinese Spring' (CS), and that of Aegilops kotschyi causes CMS in spelt wheat (T. spelta) var. duhamelianum (Sp). CS has fertility-restoring (Rf) genes against the latter cytoplasm and Sp has the ones against the former. To know the genetic system concerning to CMS, we crossed 66 F8 recombinant inbred lines (RILs) derived from a cross between CS and Sp as males to the alloplasmic lines of CS and Sp having the cytoplasms of T. timopheevi and Ae. kotschyi, respectively. The fertilities of respective F1 plants derived from the crosses were examined for QTL analysis. The major QTLs detected in both systems were located on the short arm of chromosome 1B. One minor QTL on chromosome 2B was also commonly detected in both of the systems, while other minor QTLs against T timopheevi cytoplasm were distributed on the chromosomes 2A, 4B, and 6A.     

几类异质1BL／1RS小麦雄性不育系诱导孤雌生殖性的研究

系统调查了4种异质（即偏凸,粘果,易变和二角山羊草细胞质）1BL/1RS小麦雄性不育系与其一系列异质同核系,同质异核系,异质异核系杂交,回交世代诱导孤雌生殖性的遗传变异规律,染色体分裂行为和对外表现特点,结果表明：1、异源细胞质与小麦1BL/1RS核型专一互作,有着消弱同源染色体配对,提高中期单价体细胞率的作用;2、特定细胞质背景下,专一核型内细胞单价体频率高低与诱导孤雌生殖性的频率直接相关;3、1BL/1RS易位染色体中易位片所存在系列差异以及1BL/1RS易位染色体外基因型背景不同,诱导孤雌生殖性的频率明显不同;4、提高或抑制不同杂交,回交世代间孤雌生殖频率,不育系母本或不同基因型父本有着同等重要的作用。选择最佳父母本组合杂交可明显提高或降低后代群体中的孤雌生殖性频率。     

异细胞质八倍体小堰麦(Trititrigia 8x)的选育及其性状与细胞遗传

本实验用普通小麦"中国春"(Triticum aestivum CV chinese spring)和八种异细胞质"中国春"(Aegilops vavilovii)CS、(Ae.juvenalis)CS、(Ae.crassa)CS、(Ae.comosa)CS、(Ae.unianstata)CS、(Ae.speltoides.M.)CS、(Ae.kotschyi)CS.(T.timopheevi)CS分别与八倍体小偃麦(Trititrigia 8x)"远中2"、"远中3"、"远中4"、"远中5"杂交与回交,选育出8种异细胞质"远中2"、"远中2"远中4"、"远中5"共32个类型。并对不同的异细质"中国春"小麦与八倍体小偃麦杂交当代结实率、种子萌发率、F1植株与回交后代的性状、育性、减数分裂行为进行了观察。结果表明:因细胞质类型不同杂交当代的结实率有差异,F1和回交后代育性差异较大,这种差异不仅与细胞质类型有关,而且与核亲本类型有关,表明明显的核质工作。不同的异细胞质类型对F1的某些性状如生育期、分蘖数、株高、籽粒饱满度有影响。杂种F1减数分裂行为与对照相比有变化,有的影响四分体的发育。     

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.     

4个普通小麦同核异质系的遗传特征研究

培育出具有Ｄ＾２型细胞质的普通小麦核质杂种可育系Ｄ＾２－ＣＡ８０５７,来源于Ｄ＾２型细胞质变的不育系ｍｓＤ＾２－ＣＡ８０５７和来源于普通小麦ＣＡ８０５７细胞质变异的不育系ｍｓＡ－ＣＡ８０５７。     

Variations in chloroplast proteins and nucleotide sequences of three chloroplast genes in Triticum and Aegilops.

Two alloplasmic wheat lines having the same common wheat nucleus but the cytoplasms of Aegilops crassa and Ae. columnaris together with the corresponding normal line (control) were used in the two-dimensional gel electrophoresis of soluble and thylakoid membrane proteins of the chloroplast. Three chloroplast polypeptides: the Rubisco large subunit, the beta subunit of ATP synthase, and an unidentified 31 kDa protein, differed in the common wheat and two Aegilops cytoplasms. Three chloroplast genes, atpB, atpE and trnM, that respectively encode the beta and epsilon subunits of ATP synthase and tRNA(met), were sequenced. The atpB gene differed by two synonymous base substitutions, whereas the other two genes were identical in the two Aegilops cytoplasms. From the predicted amino acid sequences, the beta subunits of the ATP synthase in the Aegilops cytoplasms were assumed to have three amino acid substitutions: Ala by Val, Asp- by Ala, and Gln by Lys+, in contrast to the cytoplasm of common wheat. This accounts for the difference in pI values found for the common wheat and Aegilops cytoplasms. The two base substitutions for the atpE genes of common wheat and the Aegilops cytoplasms were synonymous. The differences detected in the genes encoding the two subunits of ATP synthase do not appear to be ascribable to the differences in phenotypic effects for the common wheat and Aegilops cytoplasms. The base substitution rate of the atpB-atpE-trnM gene cluster was similar to that of the rbcL gene. From the rate for the atpB gene alone, evolutionary divergence of the wheat-Aegilops complex is assumed to have begun ca. 3.0 x 10(6) years ago, as compared to ca. 8.0 x 10(6) years ago for the divergence of the wheat-Aegilops complex and barley.