普通小麦不同优势杂交种及其亲本苗期根系基因的差异表达

为探讨小麦(Triticum aestivum L.)杂种优势形成的分子机理,选用普通小麦品种(系)3338、6554和2410TD及其强优势杂种A(3338×6654)和无优势杂种B(2410TD×6554),采用mRNA差异显示技术,对生长至三叶一心的根系(初生根)基因表达差异进行了比较研究.结果发现,小麦杂种一代苗期根系基因表达较亲本明显不同,表现为数量水平和质量水平上的差异,且差异表达基因的数目远高于我们以苗期叶片为材料的研究结果,表明小麦杂交种与其亲本间的基因差异表达与所研究的组织和器官有关.比较分析发现,在强优势杂种组合A中,超亲表达和偏高亲表达基因所占比例均明显高于无优势杂种组合B.以家族特异基因替代随机引物进行的差异显示结果表明,MADS-box家族基因在小麦杂交种和亲本苗期根系中存在着显著的表达差异,且差异表达类型以杂种特异表达和亲本基因在杂种一代沉默为主,说明MADS-box家族基因可能与小麦的杂种优势形成具有重要关系.对杂种和亲本基因表达差异与杂种优势的关系进行了分析和讨论.     

Identification of differentially expressed proteins between hybrid and parents in wheat (Triticum aestivum L.) seedling leaves

In spite of commercial use of heterosis in agriculture, the molecular basis of heterosis is poorly understood. To gain a better understanding of the molecular basis of wheat heterosis, we carried out a comparative proteomic analysis in seedling leaves between wheat hybrid and parents. Common wheat (Triticum aestivum L., 2n = 6x = 42, AABBDD) Line 3338 and spelt wheat (Triticum spelta L., 2n = 6x = 42, AABBDD) Line 2463 were used to produce a heterotic F₁ hybrid. The expression patterns of the total proteins were compared in seedling leaves between hybrid and its parents by using two-dimensional gel electrophoresis with two pH ranges for the first dimension separation. Among ~900 protein spots reproducibly detected, 49 protein spots were identified as being differentially expressed between hybrid and its parental lines (P < 0.05) for more than 1.5-folds. Six possible modes of differential expression were observed, including high- and low-parent dominance, underdominance, and overdominance, uniparent silencing and uniparent dominance. Moreover, 30 of the 49 differentially expressed protein spots were identified, which were involved in metabolism, signal transduction, energy, cell growth and division, disease and defense, secondary metabolism. These results indicated that wheat hybridization can cause protein expression differences between hybrid and its parents; these proteins were involved in diverse physiological process pathways, which might be responsible for the observed heterosis. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. X. Song and Z. Ni have equally contributed to this work.     

普通小麦×根茎冰草×黑麦三属杂种自交可育性的细胞学机理

普通小麦（ＴｒｉｔｉｃｕｍａｅｔｉｖｕｍＬ．,２ｎ＝６ｘ＝４２;ＡＡＢＢＤＤ）和根茎冰草（ＡｇｒｏｐｙｒｏｎｍｉｃｈｎｏｉＲｅｓｈｅｖ．,２ｎ＝４ｘ＝２８;ＰＰＰＰ）间的Ｆ１杂种（２ｎ＝５ｘ＝３５：ＡＢＤＰＰ）与两个黑麦（ＳｅｃａｌｅｃｅｒｅａｌｅＬ．,２ｎ＝２ｘ＝１４;ＲＲ）品种杂交,产生了自交可育的三属杂种。经细胞学研究表明,这种自交可育性是由于在某些细胞中通过两种方式发生了第一次减数分裂的失败,即单价染色体在赤道板发生分裂和单价染色体在一极的聚集,从而异致了有功能的雌、雄配子的形成。有功能配子的形成受染色体配对频率、基因型和环境状况的影响。冰草属的Ｐ染色体组存在染色体分离的控制基因,从而引起含有冰草属的杂种能够形成有功能的配子且自交可育。     

Production,morphology, and cytogenetics of Triticum aestivum (L.) Thell × Elymus scabrus (R. Br.) Love intergeneric hybrids obtained by in ovulo embryo culture

Summary Intergeneric hybrids were produced between common wheat, Triticum aestivum (2n=6x=42, AABBDD), and an apomictic Triticeae species, Elymus scabrus (syn. Agropyron scabrum) (2n=6x=42, HHSSSS), the first successful report of this cross. Nine tiny, underdeveloped, and structureless embryos were obtained in vitro only by in ovulo embryo culture at 4 days after pollination, which gave rise to five mature hybrid plants. All the hybrid plants were vigorous and possessed a phenotype intermediate to the two parents. There were 2n=6x=42 (ABDHSS) somatic chromosomes in the hybrids. There was little or no homology between the parental genomes, as shown by an overall meiotic chromosome association of 32.83 I + 4.08 rod II + 0.21 ring II + 0.18 III + 0.02 IV. The hybrids were completely sterile and so far backcrosses to wheat parent have not been successful. Alternate approaches to induce gene transfer(s) from E. scabrus to wheat are being attempted.Agriculture Canada Contribution No. 398.     

玉米雌穗发育期基因差异表达与杂种优势的研究

杂种优势在提高粮食作物特别是玉米的产量方面具有重要的作用。然而,杂种优势的原理却仍然是一个世界性的难题。用12个玉米自交系及其按不完全双列杂交组配的33个杂交种为材料,分4个不同发育时期取杂交种及其亲本的雌穗组织,利用差异显示技术,分析杂种与亲本的基因差异表达类型及其与7个主要农艺性状的杂种表现和杂种优势的相关关系。发现1):在5种表达类型中单态表达(基因在杂交种和双亲中同时表达的类型)的数量最大,这说明杂种优势的形成不仅与基因的表达与否相关,还与大量基因的上调或下调表达相关;2):在玉米雌幼穗的发育初期杂交种与双亲的基因表达差异最大,这可能与雌穗发育初期器官的形成和发育相关,因此这一时期差异表达(在质的方面)的基因对产量性状和杂种优势的形成具有密切关系;3):综合各种基因表达类型与产量性状和杂种优势的关系,发现某些基因在杂种中的沉默表达可以促进籽粒的发育和抑制幼穗中小花的发育。     

小麦杂交种与亲本发育早期种子的基因表达差异及其与杂种优势关系的初步研究

为深入探讨小麦杂种优势形成的分子机理,选取3个冬小麦品种(系)为一组亲本,3个为另一组亲本,配制了正反交18个杂交组合,以授粉后6d的杂交和自交种子为材料,应用mRNA差异显示技术(DDRT—PCR)研究了小麦杂交当代种子与其亲本自交种子基因的表达差异,并与杂种优势进行相关分析。为降低DDRT—PCR技术假阳性的不利影响,对每个引物组合均作了两次PCR扩增,在处理数据时,仅统计能重复出现的条带。结果发现：杂交种和亲本之间的基因表达模式有8类共15种：(1)单亲沉默型(2种),(2)单亲一致型(2种),(3)正交或反交沉默型(2种),(4)正交或反交特异型(2种),(5)正交或反交单亲一致型(4种),(6)杂交种特异型(1种),(7)双亲共沉默型(1种),(8)表达一致型(1种)。分析发现,小麦杂交种和亲本间存在显著的表达差异。在差异表达类型中,杂交种特异型和双亲共沉默型比例最低。对上述15种表达模式与杂种优势进行相关分析,结果表明,表达一致型与各产量性状杂种优势之间的相关均不显著,说明杂种优势是由某些有表达差异的基因造成。9个产量性状均能检测到一种以上与其显著或极显著相关的基因表达模式,有些性状受正负相关效应的共同影响;沉默型(包括单亲沉默型、正交或反交沉默型和双亲共沉默型)和正交或反交单亲一致型在杂种优势形成中发挥重要作用。这些研究表明,在种子发育早期,基因的差异表达与杂种优势形成之间可能存在较为复杂的关系。     

Elymus与普通小麦属间杂种的细胞遗传学研究

本研究以Elymus pendulinus(Nevski)Tzvelev(2n=4x=28,SSYY)、E.shandongenisis B.Salomon(2n=4x=28,SSYY)与普通小麦(Triticum aestivum L.:2n=6x=42,AABBDD)进行了属间远缘杂交。通过对杂种胚的离体培养,两个组合均产生了杂种F_1植株。杂种F_1为多年生,植株生长旺盛;形态上介于亲本种之间而兼具双亲的某些特征;穗状花序发育正常,但均完全不育。两个组合的根尖和花粉母细胞染色体数目为2n-5x=35。通过对杂种减数分裂染色体配对行为的分析,发现其MI染色体的配对水平很低,二价体均为棒状,每细胞的平均染色体交叉数在0.25-0.31之间。这表明E.pendulinus t E.shandongensis 所含的SY染色体组与普通小麦的ABD染色体之间的同源程度很低。由于在E.shandongeasis 及其它具有SY染色体组的Elymus 单倍体中,SY染色体组之间的部份同源染色体配对数均明显高于该杂种中的配对数,这表明存在于普通小麦中的ph基因及其它具类似作用的基因系统能抑制SY染色体组之间的部份同源染色体配对。     

A novel intergeneric hybrid in the Triticeae: Triticum aestivum x Psathyrost achys juncea

Summary Two hybrid embryos of intergeneric origin between Triticum aestivum cv Fukuho (2n=6x=42, AABBDD) and Psathyrostachys juncea (2n=2x=14, NN) were successfully rescued. One hybrid plant had the expected chromosome number of 28 (ABDN), whereas the second plant had 35 chromosomes. The average meiotic chromosome pairing in the 35-chromosome hybrid was 21.87 univalents + 6.38 bivalents + 0.11 trivalents + 0.009 quadrivalents, which indicates that two copies of the N genome were present. Chromosome pairing in the 28-chromosome hybrid was low (1.35 bivalents), and pointed out the lack of homology between the wheat genomes and the P. juncea genome. These new hybrids showed some necrosis and chlorosis, which caused severe floral abortion in the plant that had 35 chromosomes. These problems became gradually less severe after 18 months.Contrib. no. 372     

Cytological Studies on F1 Hybrids Between Hexaploid Aegilops and Triticum aestivum

Morphologcal and cytological studies of hybrids between hexaploid Aegilops crassa Boiss. (2n = 6x = 42, DDD2 D2Mcr Mcr), Ae. vavilovii (Zhuk.) Chen. (2n = 6x = 42, DDMcr McrSp Sp) and Triticum aestivum L. (2n= 6x = 42, AABBDD) were carried out. The results showed that most of the F1 hybrids morphologically resembled their Aegilops parental species. Four Fl hybrids of Ae. vavilovii × T. aestivum and one of Ae. crassa × T. aestivum produced seeds containing few endosperms. The percentage of seed obtained ranged from 0.1% to 6.5 %. These seeds were not vigorous and only a few of them germinated. A large number of univalents appeared at meiosis MI. The frequencies of bivalents were lower than those being theoriticaly estimated. These results indicated that the D genomes in Ae. Crassa and Ae. vavilovii may have been substantially modified. Trivalents were observed in all Fl hybrids. Quadrivalents and pentavalents were also observed in some PMCs during meiosis. The chiasmata frequencies in hybrids using Aegilops species as maternal parents were higher than those in their reciprocal ones. Chromosome segrigations were abnormal at A Ⅰ and A Ⅱ . Polyads and micronuclei appeared frequently at telophase tetrad stage. A plant with 21 chromosomes was obtained in Ae. vavilovii × Jimai 30, although the reason was not yet clear.     

Heterosis in root development and differential gene expression between hybrids and their parental inbreds in wheat (Triticum aestivum L.)

In spite of commercial use of heterosis in agriculture, the molecular basis of heterosis is poorly understood. In this study, heterosis was estimated for eight root traits in 20 wheat hybrids derived from a NC Design II mating scheme. Positive mid-parent heterosis was detected in 96 of 160 hybrid–trait combinations, and positive high-parent heterosis was detected in 79 of 160 hybrid-trait combinations. Improved differential display was used to analyze alterations in gene expression between hybrids and their parents in roots at the jointing stage. More than 990 fragments were repeatedly displayed, among which 27.52% were differentially expressed between hybrids and their parents. Four differential expression patterns were observed. Thirty differentially expressed cDNA fragments and three genes with open reading frames were cloned, and their expression patterns were confirmed by reverse-northern blot and semi-quantitative RT-PCR analysis, respectively. We concluded that these differentially expressed genes, though mostly with unknown function, could play important roles for hybrids to demonstrate heterosis in root system traits.Electronic Supplementary Material Supplementary material is available to authorised users in the online version of this article at .Z. Wang and Z. Ni contributed to this article equally.     

基因差异表达与杂种优势形成机制探讨

对杂种优势这一普遍而重要的生物学现象研究虽有百余年的历史, 但其根本机理尚未阐述清楚。继基因组组成差异及基因效应研究之后, 基因表达差异成为探寻杂种优势分子机理新的切入点。旨在通过揭示杂种中等位基因差异表达、杂种与亲本间基因差异表达的调控机制, 来认识杂种优势形成的分子机理, 从而达到指导育种实践的目的。文章概述了杂种等位基因差异表达现象及其产生机理, 总结了杂种与亲本相比所呈现出的加性、显性和超显性等多种差异基因表达模式, 归纳了表达谱研究筛选出的与杂种优势形成有关的基因, 以及某些关键生化代谢途径对杂种优势形成的贡献。但由于杂种优势机理的复杂性, 基因表达研究并没有得出统一的表达模式, 大多数杂种优势基因也不能被归属为同一类别。尽管如此, 基因表达谱研究毕竟迈出了解析杂种优势形成复杂基因表达网络的第一步, 随着表达谱技术和生物信息学的不断更新和发展, 杂种优势形成的分子机理有望在基因表达层面上取得突破。     

Expression of fertility during morphogenesis in self-pollinated backcrossed progenies of barley-wheat amphiploids

The fertility characteristics expressed during morphogenesis in first-generation self-pollinated backcrossed progenies (BC1) obtained from amphiploid barley-wheat hybrids [Hordeum geniculatum All. (2n = 28) x Triticum aestivum L. (2n = 42)] (2n = 70) backcrossed with common wheat were studied. It was found that, in the case of self-pollination of BC1 plants, karyotype stabilization leads to the formation of alloplasmic euploid (2n = 42), telocentric substitution (2n = 40 + 2t), and telocentric addition (2n = 42 + 2t), (2n = 42 + 2t) plant forms, which may serve as the sources of the respective alloplasmic lines of common wheat. That the expression of fertility characters in BC1F8 plants was shown to depend on growth conditions. The main mechanism of hybrid incompatibility of BC1F1-BC1F8 plants was expressed as grass-clump dwarfism.     

小麦杂种及其亲本苗期叶片家族基因差异表达及其与杂种优势关系的初步研究

为探讨小麦杂种优势形成的分子机理,以一套双列杂交组合的苗期叶片为材料,利用mRNA差异显示技术分析了杂种及其亲本间MADS－box、G－ box、Ser/Thr蛋白激酶、EIF－4A、ARF1基因家族共5类家族基因在杂交种和亲本之间的表达差异。并与杂种性状表现和杂种优势进行了相关分析。结果发现,除ARF1家族基因外,其余家族基因在杂种和亲本间存在显著的表达差异,差异表达类型可概括为4种：（1）双亲共沉默;（20单亲表达沉默;（3）杂种特异表达;（4）单亲表达一致。分析发现,MADS－box、G－box和EIF－4A家族基因在杂种和亲本间的差异表达模式相似,均以单亲特异表达和种特异表达类型所占比例最高。相关分析结果表明,以上所有家族基因的总体差异表达程度与所有性状的杂种表现均不相关,MADS－box家族基因中杂种特异表达类型与小穗数、单株产量和单穗产量杂种优势呈显著正相关,双亲共沉默类型与小穗数、千粒重和单穗产量杂种优势呈显著负相关,另外,EIF－4A家族基因中单亲表达一致型与单穗产量杂种优势呈显著正相,但双亲共沉默类型与小穗数和单穗产量杂种优势呈显著负相关,对于G－box基因家族而言,仅小穗数杂种优势和双亲共沉默类型成显著负相关,而蛋白激酶家族基因的各种差异类型与性状杂种优势的相关分析均不显著。这些研究表明,调控基因的差异表达与杂种优势形成有密切关系。     

向普通小麦导入纤毛鹅观草抗黄矮病基因的研究──Ⅰ.F_1和BC_1的产生及其细胞遗传学

为了将纤毛鹅观草Z1010对黄矮病毒株系PAV和RPV的抗性基因转入普通小麦,通过幼胚拯救,获得了纤毛鹅观草Z1010×普通小麦品种莱州953的杂种F1,以及用5个普通小麦品种（系）回交的BC1衍生系。对杂种F1及BC1植株的细胞学分析表明,纤毛鹅观草Z1010不仅对Ph基因具有很强的抑制作用,而且能使杂种F1形成未减数配子,对细胞遗传学资料的进一步分析认为,通过部分同源染色体间的交换将纤毛鹅观草Z1010的抗黄矮病基因转入小麦是可能的。     

Hybrids and backcross progenies between wheat (Triticum aestivum L.) and apomictic Australian wheatgrass [Elymus rectisetus (Nees in Lehm.) A. Löve & Connor]: karyotypic and genomic analyses

Wheat (Triticum aestivum L., 2n = 6x = 42, AABBDD) florets were emasculated and pollinated using two apomictic wheatgrass [Elymus rectisetus (Nees in Lehm.) A. Love & Connor, 2n = 6x = 42, SSYYWW] accessions, one of which produces 2n pollen. A 2n = 42 (B_II) hybrid and four 2n = 63 (B _III) hybrids were obtained. The spike morphology of the B _II hybrid was intermediate to that of its parents. The pollen mother cells (PMCs) of this hybrid contained on average 38.361 and 1.62 II, which was consistent with its disparate genome composition (ABDSYW). Its pollen failed to stain and no BC₁ progeny was obtained. The B _III hybrids (reduced egg fertilized with unreduced sperm) were grasslike and had a full complement of E. rectisetus chromosomes, the synapsis of which was slightly impaired by wheat haplome and/or cytoplasm. Their PMCs contained on average 16.30 II, 25.72 I, and 1.54 multivalents (III plus IV). Pollen stainability in these hybrids was low (<1%), and when they were used as females, one 54- and 60-chromosome BC₁ were obtained. A mean of 13.25 II was observed in PMCs of the 54-chromosome BC₁ and pollen stainability was 10%. Pollen stainability in the 60-chromosome BC₁ was only 5%. The use of 2n-pollen-producing E. rectisetus accession accelerated hybrid and BC₁ formation and may accelerate the ultimate transfer of apomixis to wheat.