拟南芥角果发育和开裂的调控机制及在油菜育种中的应用

十字花科芸苔属甘蓝型油菜(Brassica napus)在角果发育成熟过程中,因角果开裂种子散落引起产量损失最低达20%左右,最高可达50%左右。模式植物拟南芥在角果形态和开裂机制方面与甘蓝型油菜具有相似性,角果开裂主要与果瓣、胎座框和果瓣边缘层的细胞发育有关。通过对拟南芥角果发育期间基因对果瓣、胎座框和果瓣边缘层的识别和发育进行综述,明确了拟南芥角果发育和开裂的基因调控机制,提出了从模式植物到大田作物甘蓝型油菜抗裂角育种的新策略。     

Dehiscence-related expression of an Arabidopsis thaliana gene encoding a polygalacturonase in transgenic plants of Brassica napus

Pod dehiscence in Arabidopsis thaliana is accompanied by an increase in the expression of a polygalacturonase (PG). The gene encoding this mRNA has been characterized and shown to have extensive homology to a similar PG gene from Brassica napus . The A. thaliana PG promoter was fused to β -glucuronidase (GUS) and the expression of this reporter gene analysed in transgenic B. napus plants. The GUS activity was detected throughout the dehiscence zone of pods from 35 d after anthesis and expression was restricted to those cells that undergo cell separation. Expression was also detectable at the junction between the seed and the funicular tissue and in mature anthers of flowers. Transgenic plants containing the PG promoter fused to barnase were sterile as a consequence of the anthers failing to undergo dehiscence. Fertilization of PG-barnase plants resulted in the development of pods that exhibited a reduced capacity to shatter. The role of PG during cell separation processes in plants is discussed.     

Effects of gene expression on molecular evolution in Arabidopsis thaliana and Arabidopsis lyrata

We analyzed the complete genome sequence of Arabidopsis thaliana and sequence data from 83 genes in the outcrossing A. lyrata, to better understand the role of gene expression on the strength of natural selection on synonymous and replacement sites in Arabidopsis. From data on tRNA gene abundance, we find a good concordance between codon preferences and the relative abundance of isoaccepting tRNAs in the complete A. thaliana genome, consistent with models of translational selection. Both EST-based and new quantitative measures of gene expression (MPSS) suggest that codon preferences derived from information on tRNA abundance are more strongly associated with gene expression than those obtained from multivariate analysis, which provides further support for the hypothesis that codon bias in Arabidopsis is under selection mediated by tRNA abundance. Consistent with previous results, analysis of protein evolution reveals a significant correlation between gene expression level and amino acid substitution rate. Analysis by MPSS estimates of gene expression suggests that this effect is primarily the result of a correlation between the number of tissues in which a gene is expressed and the rate of amino acid substitution, which indicates that the degree of tissue specialization may be an important determinant of the rate of protein evolution in Arabidopsis.     

拟南芥不定芽发生早期的数字基因表达谱分析

目前,有关不定芽发生的研究主要集中在单基因的调控方面,缺乏转录组方面的系统研究.利用RNA-seq高通量测序技术在全基因组范围内检测了不定芽发生早期的基因表达谱,共检测到2457个差异表达基因.这些基因参与了激素代谢和信号转导、愈伤组织和侧根的形成、茎顶端分生组织的发育和光合作用等过程.进一步的途径富集分析表明,不定芽发生早期苯丙氨酸代谢和苯丙胺素合成等途径相关的基因显著富集.并且苯丙氨酸可以显著抑制不定芽的发生,暗示了苯丙氨酸代谢和苯丙胺素的合成可能在不定芽发生过程起着重要的作用.     

Characterization of expressed genes in the SLL2 region of self-compatible Arabidopsis thaliana.

Self-incompatibility in Brassica species is regulated by a set of S-locus genes: SLG, SRK, and SP11/SCR. In the vicinity of the S-locus genes, several expressed genes, SLL2 and SP2/ClpP, etc., were identified in B. campestris. Arabidopsis thaliana is a self-compatible Brassica relative, and its complete genome has been sequenced. From comparison of the genomic sequences between B. campestris and A. thaliana, microsynteny between gene clusters of Arabidopsis and Brassica SLL2 regions was observed, though the S-locus genes, SLG, SRK, and SP11/SCR were not found in the region of Arabidopsis. Almost all genes predicted in this region of Arabidopsis were expressed in both vegetative and reproductive organs, suggesting that the genes in the SLL2 region might not be related to self-incompatibility. Considering the recent speculation that the S-locus genes were translocated as a single unit between Arabidopsis and Brassica, the translocation might have occurred in the region between the SLL2 and SP7 genes.     

拟南芥FRUITFULL(FUL)基因的表达调控模式

FRUITFULL(FUL)基因是一类MADS box基因,在控制开花时间、花分生组织分化、茎生叶形态以及心皮和果实的发育中发挥重要作用。为了阐明FUL的表达调控模式,克隆了拟南芥Arabidopsis thaliana FUL启动子区(-2148bp~+96bp)及其第一内含子,并构建一系列启动子分段缺失表达载体及含FUL第一内含子的融合载体。并进一步构建了各顺式作用元件融合拟南芥TUBULIN和ACTIN启动子的表达载体。转基因拟南芥分析结果表明,FUL启动子的上游存在2个抑制其表达的顺式作用元件,其中一个很可能与转录因子AP1的结合有关;2个存在于上游调控区的CArG-box对FUL基因表达起到重要的调控作用;FUL基因第一内含子参与拟南芥心皮和雄蕊的发育调控,而且有增强基因表达的作用。     

Chromosomal location and expression of the single-copy gene encoding high-mobility-group protein HMG-I/Y in Arabidopsis thaliana

A cDNA encoding the HMG-I/Y protein from Arabidopsis thaliana has been isolated and characterised by nucleotide sequencing. The 903 bp cDNA contains a 612 bp open reading frame encoding a protein of 204 amino acid residues showing homology to HMG-I/Y proteins from other plant species. The protein contains four copies of the AT-hook motif which is involved in binding A/T-rich DNA. Southern blotting showed that the HMG-I/Y gene was present in a single copy in the Arabidopsis genome. The gene was localised to the top of chromosome 1 by RFLP analysis of F₈ recombinant inbred lines. Northern blotting showed that the gene was expressed in all organs examined, with the highest expression in flowers and developing siliques.     

拟南芥prr5突变体对ABA的响应

以拟南芥为材料,统计PRRs （pseudo-response regulators）突变体 prr5及其野生型经ABA处理后的萌发率、根长和NaCl处理后的萌发率,并采用实时定量PCR方法,对不同浓度ABA处理的拟南芥幼苗中的PRR5基因表达进行分析.结果表明：prr5突变体对ABA弱敏感,其种子萌发率比野生型显著或极显著增高,主根比野生型长,且PRR5基因表达受ABA抑制.同时,NaCl处理后,prr5的萌发率比野生型极显著增高.因此,推测prr5可能为ABA信号通路相关基因.     

Characterization and genetic mapping of a mutation (ms35) which prevents anther dehiscence in Arabidopsis thaliana by affecting secondary wall thickening in the endothecium

The male sterile mutant, ms35 , of Arabidopsis thaliana was produced by X-irradiation of seeds. The mutant produces fertile pollen, but is male sterile because the anthers do not dehisce. Anther development in ms35 plants occurs as in wild-type Arabidopsis until shortly after microspores are released from meiotic tetrads. Thereafter, in the wild type, bands of lignified, cellulosic secondary wall thickenings are laid down around the cells of the anther endothecium. In contrast, wall thickenings are not formed in the endothecium of the ms35 mutant. Development of other lignified tissues, for example the vascular tissue of the stamen, occurs normally in ms35 plants. In mutant anthers, as pollen maturation is completed, the stomium is cleaved but the anther wall does not retract to release pollen. The block in anther dehiscence in ms35 plants is specifically correlated with the absence of endothecial wall thickenings. The ms35 mutation represents the first genetic evidence in support of the proposed role of the endothecium in anther dehiscence. The ms35 gene was mapped to the top arm of chromosome 3 ( hy2 -(4.17±2.31 cM)- ms35 -(32.14±5.45 cM)- gl1 ).     

一氧化氮在拟南芥气孔发育中的功能研究

为了研究一氧化氮(nitric oxide,NO)在气孔发育中的功能,本实验利用NO的释放剂硝普钠(SNP)处理拟南芥野生型,结果显示,SNP处理后的子叶气孔指数(SI)和%(GMC+M)都较未处理组明显升高。继续检测拟南芥内源一氧化氮升高的突变体nox1和内源一氧化氮降低的突变体noa1的气孔参数,结果显示,nox1的气孔指数(SI)和%(GMC+M)相对野生型明显提高,noa1的气孔指数(SI)和%(GMC+M)低于野生型。荧光定量PCR结果进一步显示,NO抑制了气孔发育相关基因MUTE,SCRM,SCRM2的表达。综上结果表明,NO通过调控气孔发育相关基因的表达影响气孔发育。     

拟南芥反应调节因子研究进展

反应调节因子是His_Asp磷酸转移信号传导途径的重要组分。它通过在保守的Asp残基上接受由感受器转移而来的磷酸基团对下游基因进行调控 ,以对环境刺激作出反应。在高等植物拟南芥中已经发现 1 4种反应调节因子 ,它们可分为A ,B两种亚型。在结构上 ,B亚型反应调节因子的B盒序列和较长的C末端延伸使其具有转录因子的作用 ;在表达特性上 ,A亚型反应调节因子的转录受细胞分裂素和硝酸盐诱导 ;在生化特性上 ,A亚型反应调节因子具有磷酸化酶活性 ,而B亚型反应调节因子可能是AHPs的磷酸提供者     

Growth and development of Arabidopsis thaliana seedlings in interaction with fungi isolated from stem-end rot of avocado fruits

Abstract

The influence of Phomopsis viticola, Diaporthe phaseolorum and Pseudofusicoccum stromaticum isolated from avocado stem-end rot on growth of Arabidopsis seedlings was investigated. We found responses that are specific for each fungus, whereas P. viticola and D. phaseolorum strongly repressed root growth, P. stromaticum induced growth and branching, and enhanced shoot biomass production. Detailed microscopy and structural analyses revealed that the fungi did not affect cell division in root meristems but rather influenced cell elongation and differentiation, and these responses were related to changes in auxin responsiveness. P. stromaticum strongly acidifies the growth medium and this correlated with induction of the jasmonic acid responsive gene construct LOX2:GUS in leaves, whereas P. viticola and D. phaseolorum showed much reduced acidification properties. Taken together, our results show that fungal isolates from the stem-end of avocado fruits interact with Arabidopsis plants in highly diverse and contrasting manners influencing growth, patterning and defence.