光敏色素与转录因子结合直接调控植物基因表达和发育

植物的光控发育一直是植物学中一个非常活跃的研究领域,是近该领域又取得重大突破性进展,人们通过酵母双杂交技术克隆到在体内与光敏色素相互作用的转录因子,并证实被光活化的光敏色素直接进入细胞核与转录因子结构合启动基因表达,本文就此研究进展作简要介绍。     

Phylogenetic structure in the grass family (Poaceae): evidence from the nuclear gene phytochrome B

Phylogenetic analyses of partial phytochrome B (PHYB) nuclear DNA sequences provide unambiguous resolution of evolutionary relationships within Poaceae. Analysis of PHYB nucleotides from 51 taxa representing seven traditionally recognized subfamilies clearly distinguishes three early-diverging herbaceous "bambusoid" lineages. First and most basal are Anomochloa and Streptochaeta, second is Pharus, and third is Puelia. The remaining grasses occur in two principal, highly supported clades. The first comprises bambusoid, oryzoid, and pooid genera (the BOP clade); the second comprises panicoid, arundinoid, chloridoid, and centothecoid genera (the PACC clade). The PHYB phylogeny is the first nuclear gene tree to address comprehensively phylogenetic relationships among grasses. It corroborates several inferences made from chloroplast gene trees, including the PACC clade, and the basal position of the herbaceous bamboos Anomochloa, Streptochaeta, and Pharus. However, the clear resolution of the sister group relationship among bambusoids, oryzoids, and pooids in the PHYB tree is novel; the relationship is only weakly supported in ndhF trees and is nonexistent in rbcL and plastid restriction site trees. Nuclear PHYB data support Anomochlooideae, Pharoideae, Pooideae sensu lato, Oryzoideae, Panicoideae, and Chloridoideae, and concur in the polyphyly of both Arundinoideae and Bambusoideae.     

葡萄Actin基因家族的鉴定及进化和表达分析

采用生物信息学方法从葡萄(Vitis vinifera Linn.)全基因组中鉴定Actin基因家族,并对各基因的染色体定位和结构特征,编码蛋白质的理化性质、亚细胞定位、二级结构、三级结构和系统进化,以及不同组织的基因表达进行研究.结果表明:葡萄Actin基因家族16个基因分布在12条染色体上.16个基因的结构特征及其编码蛋白质的理化性质差异较大.16个基因的长度及其内含子总长度的变化范围较大,编码序列(CDS)和外显子总长度的变化范围较小.除登录号GSVIVG01008254001和GSVIVG01014035001的基因外,其他14个基因的GC含量均低于其CDS的GC含量.除登录号GSVIVG01008254001的基因外,其他15个基因编码的蛋白质的理论相对分子质量为12534.54~82612.33,理论等电点为pI 4.92~pI 9.13.16个基因编码蛋白质的消光系数为14105~73645,脂肪族氨基酸指数为65.54~92.06,其中9个为稳定蛋白,7个为不稳定蛋白.除登录号GSVIVG01014035001的基因外,其他15个基因编码的蛋白质均为亲水性蛋白.登录号GSVIVG01016517001的基因编码的蛋白质定位于细胞质和细胞核,其他15个基因编码的蛋白质定位于细胞质.二级结构和三级结构显示:葡萄Actin基因家族16个基因编码的蛋白质均由α螺旋、无规则卷曲和延伸链构成,且总体以无规则卷曲为主.系统进化分析和不同组织的基因表达分析结果显示:与拟南芥〔Arabidopsis thaliana(Linn.)Heynh.〕相似,葡萄Actin基因家族16个基因编码的蛋白质分为3个亚家族,ClassⅡ亚家族(营养型)包括登录号GSVIVG01003099001和GSVIVG01026580001的基因编码的蛋白质,这2个基因在所有组织中的表达均较高;ClassⅢ亚家族(生殖型)包括登录号GSVIVG01033494001、GSVIVG01024980001和GSVIVG01016550001的基因编码的蛋白质,这3个基因在花粉、雄蕊和花中的表达均较高;ClassⅠ亚家族包括其他11个基因编码的蛋白质,这11个基因在各组织中的表达总体上较低.研究结果显示:葡萄Actin基因家族的表达具有组织特异性.     

Exon-intron structure and evolution of the Lipocalin gene family

The Lipocalins are an ancient protein family whose expression is currently confirmed in bacteria, protoctists, plants, arthropods, and chordates. The evolution of this protein family has been assessed previously using amino acid sequence phylogenies. In this report we use an independent set of characters derived from the gene structure (exon-intron arrangement) to infer a new lipocalin phylogeny. We also present the novel gene structure of three insect lipocalins. The position and phase of introns are well preserved among lipocalin clades when mapped onto a protein sequence alignment, suggesting the homologous nature of these introns. Because of this homology, we use the intron position and phase of 23 lipocalin genes to reconstruct a phylogeny by maximum parsimony and distance methods. These phylogenies are very similar to the phylogenies derived from protein sequence. This result is confirmed by congruence analysis, and a consensus tree shows the commonalities between the two source trees. Interestingly, the intron arrangement phylogeny shows that metazoan lipocalins have more introns than other eukaryotic lipocalins, and that intron gains have occurred in the C-termini of chordate lipocalins. We also analyze the relationship of intron arrangement and protein tertiary structure, as well as the relationship of lipocalins with members of the proposed structural superfamily of calycins. Our congruence analysis validates the gene structure data as a source of phylogenetic information and helps to further refine our hypothesis on the evolutionary history of lipocalins.     

枸杞耐盐变异体的筛选及植株再生

枸杞(Lycium barbarum L.)无菌苗下胚轴于MS+2,4-D 0.25mg/L+LH 50 0mg/L的诱导培养基上产生胚性愈伤组织。经0.34%的EMS(半致死剂量)处理并恢复增殖2周后, 将存活组织转接到含有1.5%NaCl的诱导培养基上培养4周,再将少数存活的组织转移到含1.0%NaCl的同样培养基上继续培养,经不断选择,选出了耐1.0%NaCl的愈伤组织变异体。经耐盐性、耐盐稳定性、脯氨酸含量、叶绿素含量分析,以及对山梨醇、聚乙二醇的反应证明,该愈伤组织是耐盐变异体。变异体在含有1.0%NaCl的分化培养基(MS+6-BA0.5mg/L)上可分化出再生植株。     

Structure and expression of a rice hsp70 gene

A genomic hsp70 gene was isolated from a rice IR36 genomic library and 4 794 bp of the gene have been sequenoed. The 5' flanking region of the gene contained a putative TATA box and a typical heat shock element sequence 5'-CTcgGAAccTTCgAG-3'. The amino acid sequence of the rice HSP70 deduced from the coding region shared 84%-92% homologies with those of HSP70s from other plant species. An intron 1939bp long was identified in the coding region at the codon specifying amino acid 72 (Asp), the similar position introns occurring in other intron-containing hsp70 genes. In addition, another intron of 57 bp was found in the 3'-untranslated region in the rice hsp70 gene. Southern blot hybridization showed that rice hsp70 gene family contained at least three members. Analysis of the RNA leveis with the gene-specific and non-specific probes revealed that the rice hsp70 gene expressed at normal temperature and the expression was enhanced by heat shock treatment.     

水稻胞质核糖体蛋白基因OsRPS7的克隆与序列分析

以已公布的黑麦胞质核糖体蛋白基因ScRPS7的cDNA序列为信息探针,在中国华大水稻基因组数据库中搜索与之高度同源的基因组重叠群。采用计算机拼接和RT-PCR方法克隆了水稻胞质核糖体蛋白基因的全长cDNA序列,命名为OsRPS7。该cDNA序列全长919bp,编码192个氨基酸;其与黑麦、拟南芥和芸薹的S7核糖体蛋白的氨基酸一致率分别为88%、72%和72%。对OsRPS7 的基因组结构和基因的功能进行了分析和预测。Abstract：Using the cDNA of rye cytoplasmic ribosomal protein ScRPS7 as a query probe, a highly homologous rice genomic contig was obtained from Huada rice genome database. The full-length cDNA sequence of rice cytoplasmic ribosomal protein S7 was assembled by informatics based on the contig. Furthermore, with the two primers designed according to this assembled cDNA, the full-length cDNA of rice ribosomal protein was cloned by RT-PCR and named as OsRPS7. The cDNA was 919bp in length and contained a complete Open Reading Frame (ORF) of 576bp, encoding a protein of 192 amino acid residues. The deduced amino acids of OsRPS7 showed 88%、72% and 72% identity with those from Secale cereale、Arabidopsis thaliana and Brassica oleracea, respectively. The genome structure of OsRPS7 was analyzed, and its function was predicted in this paper.     

Genomic structure,expression and evolution of the alfalfa aspartate aminotransferase genes

Genomic clones encoding two isozymes of aspartate aminotransferase (AAT) were isolated from an alfalfa genomic library and their DNA sequences were determined. The AAT1 gene contains 12 exons that encode a cytosolic protein expressed at similar levels in roots, stems and nodules. In nodules, the amount of AAT1 mRNA was similar at all stages of development, and was slightly reduced in nodules incapable of fixing nitrogen. The AAT1 mRNA is polyadenylated at multiple sites differing by more than 250 bp. The AAT2 gene contains 11 exons, with 5 introns located in positions identical to those found in animal AAT genes, and encodes a plastid-localized isozyme. The AAT2 mRNA is polyadenylated at a very limited range of sites. The transit peptide of AAT2 is encoded by the first two and part of the third exon. AAT2 mRNA is much more abundant in nodules than in other organs, and increases dramatically during the course of nodule development. Unlike AAT1, expression of AAT2 is significantly reduced in nodules incapable of fixing nitrogen. Phylogenetic analysis of deduced AAT proteins revealed 4 separate but related groups of AAT proteins; the animal cytosolic AATs, the plant cytosolic AATs, the plant plastid AATs, and the mitochondrial AATs.     

Sex-related genes expression in juveniles of red abalone,Haliotis rufescens (Swanson, 1822)

The red abalone, Haliotis rufescens (Swanson 1822), is an important species for commercial aquaculture in Mexico, with annual production levels around 68 t. Not surprisingly, there is great interest in increasing production and its cultivation success. In order to have a better understanding of the genes involved in the sexual differentiation, this study analyzed the early differential expression of eight sex-related genes (VCP 2.2, VERL, VTGI, DMRT1, FP, LYS, SARIP, TEKT A1) by RT-qPCR in abalones from 5 to 45 mm of shell length. Sex identification was evaluated using VERL and LYS genes expression levels. These genes differentiated females and males from 16 mm of shell length and above. All eight sex-specific genes showed expression in all organisms. However, their level was higher in the sex group to which they correspond. In contrast, differential expression levels occurred in individuals with a shell length of 26–35 mm and 36–45 mm. These results suggest that sex gene expression is not entirely sex specific, and sexual differentiation in red abalone occurs between 16 and 25 mm of shell length. More studies are needed to determine the function of these genes in the sex that they are not associated with.