葡萄NIP基因家族的鉴定与表达分析

类根瘤菌26膜内在蛋白(nodulin 26 like intrinsic proteins,NIPs)是水通道蛋白的亚类,在植物营养获取和胁迫应答过程中发挥着重要作用。该研究利用多种生物信息学软件,对葡萄NIP家族基因进行分析,并采用RT PCR方法克隆得到4个NIP家族基因,利用qRT PCR方法分析非生物胁迫下NIP基因的表达特征。结果显示：(1)在葡萄基因组中,共鉴定到8个NIP基因,分布于葡萄4条染色体上,主要定位在质膜中;结构上含有6个跨膜结构域和两个典型的保守结构域NPA;氨基酸序列中存在很多个可能的磷酸化位点。(2)进化分析表明葡萄和拟南芥NIP基因具有较高的同源性,基因结构包含外显子数4~6个,保守基序种类和数量相似;基因启动子上游2 kb包含多种应答逆境和激素的顺式调控元件,其数量差异可能与基因本身功能相关。(3)NIP家族基因在不同组织中表达水平差异较大,多数成员在叶中表达水平较高,在茎中较低;成功克隆得到4个葡萄VvNIP基因,其长度分别为789 bp、606 bp、897 bp、789 bp,分别编码262、201、298、293个氨基酸。(4)qRT PCR结果显示,不同胁迫处理下NIP基因在葡萄叶片中的表达水平不同：低温处理下葡萄NIP基因大多呈显著下调表达;盐胁迫下,除VvNIP2 1、VvNIP4 2外其余家族基因均呈下调表达;干旱胁迫下VvNIP4 2显著上调。研究表明,VvNIP基因对多种胁迫均有响应,为葡萄逆境胁迫机制研究提供了参考。     

谷子CPP基因家族鉴定及外源硒响应分析

【目的】探究谷子CPP家族成员基本特征及其在外源硒处理下的响应模式,为谷子富硒高叶酸新品种选育提供新的遗传材料。【方法】借助生物信息学工具鉴定谷子CPP家族成员,采用qRT-PCR技术对CPP基因在不同组织及外源硒处理下的表达水平进行检测,为揭示谷子CPP家族生物学功能及外源硒响应机制奠定基础。【结果】(1)谷子基因组中包含9个CPP基因,定位于6条染色体上,依照基因在染色体的排布命名为SiCPP1～SiCPP9,氨基酸数目在220～899之间,相对分子量为23624.32～97253.16Da,等电点介于6.64～9.17之间,内含子数目为3～11个不等,亚细胞预测显示该家族所有成员均能在细胞核定位到,蛋白质二级结构预测表明该家族中占比最大的是α-螺旋其次是无规则卷曲。(2)系统进化分析表明,谷子CPP蛋白可分为4个亚家族,相同亚家族之间保守基序和结构域的数目及分布相似。(3)启动子分析发现谷子CPP家族中存在大量光、生长发育、激素以及胁迫响应元件。(4)荧光定量结果显示,谷子CPP家族成员在根、茎、叶和穗中差异表达。谷子CPP家族9个成员均对外源硒存在响应,其中SiCPP5、SiCPP6、SiCPP7和SiCPP8响应程度最强。【结论】谷子CPP家族成员具有组织表达特异性且对外源硒存在不同程度响应。     

辣椒锌指蛋白DnaJ-Like基因家族鉴定及对高温胁迫的表达响应

植物DnaJ-Like锌指蛋白（DNAJE）是近年来发现的一类具有重要功能的蛋白,在光合作用、质体发育与运动和植物抗逆及防御反应中发挥重要作用。基于辣椒全基因组数据,对辣椒DNAJE基因家族（CaDNAJE）进行鉴定,利用生物信息学方法对基因和蛋白特征、比较进化、共线性关系及高温胁迫下基因表达模式等进行分析。结果表明,CaDNAJE基因家族有28个成员,不均匀的分布在10条染色体和3个Scaffold上,氨基酸序列长度为99-470 aa,分子量为10.05-52.26 kD,理论等电点（pI）为4.75-9.64,其编码蛋白主要定位在叶绿体和细胞核中;辣椒、番茄和拟南芥的比较进化分析可将其分为GroupⅠ 和GroupⅡ 2个亚族,同一亚族具有类似的蛋白保守motif和基因结构;CaDNAJE基因启动子区包含大量光响应、激素响应和胁迫应答元件;辣椒与拟南芥DNAJE之间存在10对共线性基因,辣椒种内仅有1对;辣椒不同组织转录组分析发现,GroupⅠ 中多数成员在各组织中均有高表达,而GroupⅡ 中除CaDNAJE2、CaDNAJE9等基因存在高表达外,多数基因表达量都很低甚至不表达;高温胁迫下,辣椒叶片过氧化氢含量急速上升后降低,丙二醛含量持续升高,抗氧化物酶活性也有不同程度的提高。同时,高温诱导热激转录因子Hsf和热激蛋白HSP耐热相关基因及CaDNAJE基因高表达,在不同胁迫时期发挥调控作用。推测CaDNAJE基因家族可能参与辣椒响应高温胁迫,提高其耐受力,以降低细胞损伤。     

Limitations to the Accumulation of Starch in the Developing Wheat Grain

The postulate that photosynthetic capacity limits the rate ofaccumulation of starch and dry matter in the developing wheatgrain has been tested by defoliating plants growing in the field,and by culturing detached ears on solutions of sucrose. In mostcases complete defoliation did not reduce the amounts of starchor dry matter produced 4 to 10 days later, or affect the amountsof sucrose in the grain. Increasing the potential supply ofsucrose above normal levels by culturing did not lead to increasesin either the deposition of starch or the quantity of sucrosein the grain. Removing leaves caused small depressions in thegain in fresh weight of the grain, and where defoliation didreduce the accumulation of starch there was no comparable effecton the amount of sucrose in the grain. Accordingly, it is concludedthat the postulate stated above is invalid. The case in support of the concept that it is the mechanismregulating the passage of sucrose into the grain which imposesa limitation on the accumulation of starch is presented.     

Genome- Wide Analysis and Characterization of the TRX Gene Family in Upland Cotton

Thioredoxins (TRX) are small molecules of proteins that are present in all organisms. TRXs play an important role in diverse functions of plant growth and development. In this study, we performed genome-wide, characterization and expression levels of TRX gene family in cotton. A total of 150 GhTRX proteins were identified in upland cotton and classified into five subfamilies based on their domain compositions. Phylogenetic tree analysis divided TRX genes into seven subgroups. GhTRX genes covered all upland cotton chromosomes, with duplicated gene events. Ka/Ks ratio of three gene pairs was less than 1, suggesting purifying selection. The functions of GhTRX genes were studied using gene ontology, protein localization, and promoter analysis. Furthermore, six GhTRX genes were randomly selected to examine their expression level in cotton development and under various exogenous treatments. The genes showed high expressions in various tissues and at different stages of leaf senescence, also showed high expression under abscisic acid, ethylene, drought, and salinity. This study reveals the first report of TRX family genes in upland cotton. However further studies are needed to elucidate their specific functions in cotton plant.     

Evolution of the X-linked Zinc Finger Gene and the Y-linked Zinc Finger Gene in Primates

We have sequenced the partial exon of the zinc finger genes (ZFX and ZFY) in 5 hominoids, 2 Old World monkeys, 1 New World monkey, and 1 prosimian. Among these primate species, the percentage similarities of the nucleotide sequence of the ZFX gene were 96-100% and 91.2-99.7% for the ZFY gene. Of 397 sites in the ZFX and ZFY gene sequences, 20 for ZFX gene and 42 for ZFY gene were found to be variable. Substitution causes 1 amino acid change in ZFX, and 5 in ZFY, among 132 amino acids. The numbers of synonymous substitutions per site (Ks) between human and the chimpanzee, gorilla and orangutan for ZFY gene were 0.026, 0.033, and 0.085, respectively. In contrast, the Ks value between human and hominoid primates for the ZFX gene was 0.008 for each comparison. Comparison of the ZFX and ZFY genes revealed that the synonymous substitution levels were higher in hominoids than in other primates. The rates of synonymous substitution per site per year were higher in the ZFY exon than in the SRY exon, and higher in the ZFY exon than in the ZFY intron, in hominoid primates.     

PcG家族CBX2基因的发现及其在哺乳动物性发育中的作用

Polycomb group complex (PcG)作为发挥转录抑制作用的重要表观遗传调控复合物参与发育、衰老以及肿瘤发生等重要生理过程。Pc G家族成员,包括PRC1 (Polycomb repressive complex 1)与PRC2 (Polycomb repressive complex 2)两种复合物,各组分间功能既协同,又不失特性,色素框同源蛋白(Chromobox homolog CBX)是PRC1的主要核心蛋白,其家族的成员之一CBX2近年来成为研究的热点。本研究综述了CBX2基因的发现、基因和蛋白质的结构组成及其在人和鼠不同组织的表达情况,阐述了CBX2在哺乳动物性发育中的作用及研究进展,发现CBX2除参与PcG复合体发挥转录抑制作用外,还具有基因转录激活作用,它作为哺乳动物性别决定级联中SRY (Sex region of Y chromosome)的上游基因可直接激活靶基因SF1/NR5A1的表达,如果CBX2基因突变或缺失将使雄性小鼠发生性反转,在人类中造成46,XY性发育障碍(disorders of sex development, DSD),为进一步研究CBX2基因在动物性发育调控中的作用提供依据。     

蚕豆蛋白质亚基分析与特异种质鉴定

采用SDS-PAGE方法,对112份不同基因型蚕豆的清蛋白和球蛋白亚基的差异性进行分析。结果显示:(1)蚕豆清蛋白和球蛋白亚基的有效等位变异分别为1.750 0±0.452 3、1.545 5±0.522 2,多态性比率分别为75.00%、54.55%,清蛋白的亚基遗传多样性指数较球蛋白亚基高。(2)蚕豆清蛋白和球蛋白分别含有在不同基因型蚕豆中构成不同的12和10个亚基,其中清蛋白含有9个基本亚基,116kD、96kD、45kD为清蛋白的3个特异亚基;球蛋白含有8个基本亚基,58kD、35kD为球蛋白的2个特异亚基;研究共鉴定筛选出42个含有清蛋白特异亚基和21个含有球蛋白亚基的种质资源。(3)清蛋白的97kD、63kD基本亚基和球蛋白的97kD、56kD、47kD基本亚基存在一定的缺失现象,共鉴定出19个清蛋白亚基和21个球蛋白亚基缺失的优异种质。研究表明,蚕豆清蛋白和球蛋白亚基构成在不同种质之间具有差异性,除基本亚基外,部分种质还含有特异亚基或缺失亚基。     

香蕉几丁质酶基因家族的鉴定及在枯萎病侵染时的表达分析

几丁质酶(CHi,Chitinase)是一种能够催化真菌细胞壁中几丁质水解的病程相关蛋白,在植物与真菌互作中起着重要的作用。本研究以拟南芥几丁质酶基因的蛋白序列为查询序列,在香蕉A基因组数据库中进行BLASTp比对,鉴定出23个香蕉几丁质酶(MaCHi,Musa chitinase)基因家族成员。根据其染色体上的位置,分别命名为MaCHi01～MaCHi23。系统进化树分析表明23个MaCHis可分为糖苷水解酶18(GH18,glycosyl hydrolase 18)和糖苷水解酶19(GH19,glycosyl hydrolase 19)2个亚家族。转录组数据分析表明接种尖孢镰刀菌古巴专化型热带4号生理小种(Foc TR4,Fusarium oxysporum f. sp. cubense tropical race 4)后MaCHi05、MaCHi06、MaCHi07、MaCHi11、MaCHi19和MaCHi20基因在感病品种巴西蕉中显著下调或不表达,在抗病品种GCTCV-119中显著上调。利用RT-qPCR技术对MaCHi05、MaCHi06、MaCHi07、MaCHi11和M...     

Rice Mitogen-activated Protein Kinase Gene Family and Its Role in Biotic and Abiotic Stress Response

The mitogen-activated protein kinase (MAPK) cascade is an important signaling module that transduces extracellu-lar stimuli into intracellular responses in eukaryotic organisms. An increasing body of evidence has shown that theMAPK-mediated cellular signaling is crucial to plant growth and development, as well as biotic and abiotic stressresponses. To date, a total of 17 MAPK genes have been identified from the rice genome. Expression profiling,biochemical characterization and/or functional analysis were carried out with many members of the rice MAPKgene family, especially those associated with biotic and abiotic stress responses. In this review, the phylogeneticrelationship and classification of rice MAPK genes are discussed to facilitate a simple nomenclature and standardannotation of the rice MAPK gene family. Functional data relating to biotic and abiotic stress responses are re-viewed for each MAPK group and show that despite overlapping in functionality, there is a certain level of functionalspecificity among different rice MAP kinases, The future challenges are to functionally characterize each MAPK, toidentify their downstream substrates and upstream kinases, and to genetically manipulate the MAPK signalingpathway in rice crops for the improvement of agronomically important traits.     

Rice Mitogen-activated Protein Kinase Gene Family and Its Role in Biotic and Abiotic Stress Response

The mitogen-activated protein kinase （MAPK） cascade is an important signaling module that transduces extracellular stimuli into intracellular responses in eukaryotic organisms. An increasing body of evidence has shown that the MAPK-mediated cellular signaling is crucial to plant growth and development, as well as biotic and abiotic stress responses. To date, a total of 17 MAPK genes have been Identified from the rice genome. Expression profiling, biochemical characterization and/or functional analysis were carried out with many members of the rice MAPK gene family, especially those associated with biotic and abiotic stress responses. In this review, the phylogenetic relationship and classification of rice MAPK genes are discussed to facilitate a simple nomenclature and standard annotation of the rice MAPK gene family. Functional data relating to biotic and abiotic stress responses are reviewed for each MAPK group and show that despite overlapping in functionality, there is a certain level of functional specificity among different rice MAP kinases. The future challenges are to functionally characterize each MAPK, to identify their downstream substrates and upstream kinases, and to genetically manipulate the MAPK signaling pathway in rice crops for the Improvement of agronomically important traits.     

Identification of Genomic Regions and the Isoamylase Gene for Reduced Grain Chalkiness in Rice

Grain chalkiness is an important grain quality related to starch granules in the endosperm. A high percentage of grain chalkiness is a major problem because it diminishes grain quality in rice. Here, we report quantitative trait loci identification for grain chalkiness using high-throughput single nucleotide polymorphism genotyping of a chromosomal segment substitution line population in which each line carried one or a few introduced japonica cultivar Nipponbare segments in the genetic background of the indica cultivar ZS97. Ten quantitative trait loci regions were commonly identified for the percentage of grain chalkiness and the degree of endosperm chalkiness. The allelic effects at nine of these quantitative trait loci reduced grain chalkiness. Furthermore, a quantitative trait locus (qPGC8-2) on chromosome 8 was validated in a chromosomal segment substitution line–derived segregation population, and had a stable effect on chalkiness in a multiple-environment evaluation of the near-isogenic lines. Residing on the qPGC8-2 region, the isoamylase gene (ISA1) was preferentially expressed in the endosperm and revealed some nucleotide polymorphisms between two varieties, Nipponbare and ZS97. Transgenic lines with suppression of ISA1 by RNA interference produced grains with 20% more chalkiness than the control. The results support that the gene may underlie qPGC8-2 for grain chalkiness. The multiple-environment trials of the near-isogenic lines also show that combination of the favorable alleles such as the ISA1 gene for low chalkiness and the GS3 gene for long grains considerably improved grain quality of ZS97, which proves useful for grain quality improvement in rice breeding programs.     

甘蓝型油菜ZF-HD基因家族的鉴定与系统进化分析

ZF-HD是一类植物特有的转录因子, 在植物生长发育及胁迫响应过程中发挥重要作用。利用生物信息学方法, 在甘蓝型油菜(Brassica napus)基因组中鉴定到62个ZF-HD基因, 其中83.9%的基因缺乏内含子, 93.5%的BnZF-HD等电点大于7, 预测定位于细胞核的蛋白大多由100个以上氨基酸组成。根据进化关系可将其分为6个亚群, 在每个亚群中, 甘蓝(B. oleracea)和白菜(B. rapa)的ZF-HD基因数量相等或近似相等, 而甘蓝型油菜的ZF-HD基因数量接近或等同于甘蓝和白菜的ZF-HD基因数量之和。同一亚群的motif数量和类型高度相似。共线性分析结果显示, 全基因组三倍体化使ZF-HD基因在二倍体祖先得到扩张, 而异源多倍体化又进一步使甘蓝型油菜ZF-HD基因家族扩张。K_a/K_s值说明大多数ZF-HD基因在进化过程中受到了纯化选择。所有BnZF-HD基因都具有光响应元件, 2/3的基因具有MeJA、ABA和厌氧诱导顺式作用元件, 推测这些基因可能参与相关生物学过程。研究结果为进一步挖掘该家族基因的生物学功能奠定基础, 同时为揭示多基因家族在异源多倍体中的进化式样提供借鉴。     

甘蓝型油菜ZF-HD基因家族的鉴定与系统进化分析

ZF-HD是一类植物特有的转录因子, 在植物生长发育及胁迫响应过程中发挥重要作用。利用生物信息学方法, 在甘蓝型油菜(Brassica napus)基因组中鉴定到62个ZF-HD基因, 其中83.9%的基因缺乏内含子, 93.5%的BnZF-HD等电点大于7, 预测定位于细胞核的蛋白大多由100个以上氨基酸组成。根据进化关系可将其分为6个亚群, 在每个亚群中, 甘蓝(B. oleracea)和白菜(B. rapa)的ZF-HD基因数量相等或近似相等, 而甘蓝型油菜的ZF-HD基因数量接近或等同于甘蓝和白菜的ZF-HD基因数量之和。同一亚群的motif数量和类型高度相似。共线性分析结果显示, 全基因组三倍体化使ZF-HD基因在二倍体祖先得到扩张, 而异源多倍体化又进一步使甘蓝型油菜ZF-HD基因家族扩张。K_a/K_s值说明大多数ZF-HD基因在进化过程中受到了纯化选择。所有BnZF-HD基因都具有光响应元件, 2/3的基因具有MeJA、ABA和厌氧诱导顺式作用元件, 推测这些基因可能参与相关生物学过程。研究结果为进一步挖掘该家族基因的生物学功能奠定基础, 同时为揭示多基因家族在异源多倍体中的进化式样提供借鉴。     

Genome-Wide Identification and Analysis of the TIFY Gene Family in Grape

Background

The TIFY gene family constitutes a plant-specific group of genes with a broad range of functions. This family encodes four subfamilies of proteins, including ZML, TIFY, PPD and JASMONATE ZIM-Domain (JAZ) proteins. JAZ proteins are targets of the SCF^COI1 complex, and function as negative regulators in the JA signaling pathway. Recently, it has been reported in both Arabidopsis and rice that TIFY genes, and especially JAZ genes, may be involved in plant defense against insect feeding, wounding, pathogens and abiotic stresses. Nonetheless, knowledge concerning the specific expression patterns and evolutionary history of plant TIFY family members is limited, especially in a woody species such as grape.

Methodology/Principal Findings

A total of two TIFY, four ZML, two PPD and 11 JAZ genes were identified in the Vitis vinifera genome. Phylogenetic analysis of TIFY protein sequences from grape, Arabidopsis and rice indicated that the grape TIFY proteins are more closely related to those of Arabidopsis than those of rice. Both segmental and tandem duplication events have been major contributors to the expansion of the grape TIFY family. In addition, synteny analysis between grape and Arabidopsis demonstrated that homologues of several grape TIFY genes were found in the corresponding syntenic blocks of Arabidopsis, suggesting that these genes arose before the divergence of lineages that led to grape and Arabidopsis. Analyses of microarray and quantitative real-time RT-PCR expression data revealed that grape TIFY genes are not a major player in the defense against biotrophic pathogens or viruses. However, many of these genes were responsive to JA and ABA, but not SA or ET.

Conclusion

The genome-wide identification, evolutionary and expression analyses of grape TIFY genes should facilitate further research of this gene family and provide new insights regarding their evolutionary history and regulatory control.     

Genome-Wide Identification of SWEET Gene Family and Its Response to Abiotic Stresses in Valencia Sweet Orange

Plant Molecular Biology Reporter - The sugars-will-eventually-be-exported-transporters (SWEET) family is a newly characterized group of sugar transporters, which plays an important role in diverse...