大豆苹果酸脱氢酶基因的电子克隆与生物信息学分析

利用电子克隆技术获得大豆中的苹果酸脱氢酶基因cDNA序列,并对基因及其编码蛋白进行生物信息学分析和预测,为进一步克隆该基因、解决植物磷高效基因数量缺乏等问题提供依据。结果表明,大豆中的苹果酸脱氢酶基因(GmMDH1)cDNA序列长度为1 574 bp,开放阅读框1 230 bp,编码409个氨基酸残基;编码蛋白含有NAD bindingsite,Dimerization interface,Substrate binding site,MDH_glyoxysomal_mitochondrial结合位点和保守域,属于LDH_MDH_like Superfamily家族;亚细胞定位分析显示,编码蛋白位于植物细胞的叶绿体中。     

甜樱桃PaGAST基因的电子克隆及生物信息学分析

为获得甜樱桃PaGAST基因的cDNA序列,并预测该基因编码蛋白的结构与功能,以草莓FaGAST1基因序列为探针,通过基于NCBI数据库中表达序列标签的电子克隆技术对甜樱桃PaGAST基因进行克隆。利用生物信息学方法对其编码蛋白的理化性质、疏水性和亲水性、信号肽序列、跨膜结构域、亚细胞定位及功能等方面进行分析。结果表明:甜樱桃PaGAST基因长度为750 bp,开放阅读框长度为324 bp,编码107个氨基酸,N末端存在信号肽序列,C末端含有保守的GASA结构域。由于PaGAST蛋白中含有的疏水性氨基酸残基较多,该蛋白具有跨膜螺旋区,是一种跨膜蛋白,且有10个预测的蛋白激酶磷酸化位点。亚细胞定位分析表明,PaGAST蛋白分布在细胞膜外的可能性很大。功能预测显示,PaGAST基因可能具有响应胁迫应答、信号转导和免疫应答方面的功能。进化分析显示甜樱桃PaGAST蛋白与桃的亲缘关系最近。在一定程度上为甜樱桃PaGAST基因的克隆及功能鉴定奠定理论基础。     

枸杞LbCO基因的克隆与生物信息学分析

CONSTANS(CO)基因是高等植物叶片中光周期途径的关键成分,生物钟及光信号控制CO基因的表达。本研究以宁夏枸杞(Lycium barbarum L.)品种之一‘宁杞7号’叶片为实验材料,通过转录组测序结果筛选,利用RT-PCR技术扩增得到‘宁杞7号’的CO基因cDNA全长序列,命名为LbCO。利用生物信息学手段对所获得的序列进行结果及功能预测,结果显示:LbCO基因的cDNA序列全长1224 bp,编码407个氨基酸,分子质量为44.83 k D,理论等电点pI=5.58,不稳定指数为39.66,是一种稳定的非分泌蛋白。该蛋白亚细胞定位于细胞核,二级结构中无规则卷曲占比最高(54.48%),其次为α-螺旋(28.99%)。系统进化分析表明枸杞LbCO蛋白与其它物种的CO蛋白具有较高的同源性,其中与辣椒属CO蛋白亲缘关系最近。本研究为后续进一步探讨该基因编码的蛋白在调控枸杞花期研究方面提供理论参考。     

沙棘PEPCK基因的克隆与生物信息学分析

磷酸烯醇式丙酮酸羧激酶(PEPCK)是调节植物糖异生途径的限速酶。本研究从沙棘果实的转录组测序结果中筛选出PEPCK基因序列,利用RT-PCR技术对其进行克隆,利用生物信息学相关方法分析其编码蛋白的理化性质和结构特征。结果表明,该基因开放阅读框(open reading frame, ORF)长度为1 989 bp,编码一个由662个氨基酸组成的亲水性蛋白,主要由α-螺旋(30.21%)、β-折叠(17.22%),无规则卷曲(47.13%)和β-转角(5.44%)组成,其ORF序列与克莱门柚(Citrus clementina)同源性最高,氨基酸序列与川桑(Moras notabilis)和洋蓟(Cynara cardunculus var. scolymus)的有较高的同源性。该结果为进一步研究沙棘PEPCK蛋白功能以及揭示植物糖异生途径的调控机制提供理论基础。     

薏苡Waxy基因的克隆与生物信息学分析

薏苡Waxy基因是控制薏苡直链淀粉合成的关键基因,对其进行生物信息学分析,为深入研究该基因的作用奠定基础并为进一步改良糯性新材料提供理论支撑,以薏苡的嫩茎为实验材料,采用RT-PCR方法克隆薏苡Waxy基因,使用NCBI、DNAMAN及ExPYSy等一系列在线软件及工具,对薏苡Waxy基因的编码区,氨基酸序列及蛋白质的结构和功能进行生物信息学分析,研究表明薏苡Waxy基因有13个内含子,全长1 845 bp,编码由614个氨基酸,属于糖基转移酶超家族,薏苡与高粱亲缘关系较近,同源性达93%;该蛋白其中α螺旋占35.18%,无规则卷曲所占比例33.22%,保守性较强,通过对该蛋白保守性分析发现:糖基转移酶、糖原/淀粉合成酶和淀粉合成酶催化区3个保守性结构域。     

小麦Wcor15基因上游调控区克隆及生物信息学分析

为了克隆小麦Wcor15基因启动子序列和预测可能的表达调控途径,试验设计特异引物采用PCR方法从小麦基因组DNA中直接进行扩增并测序,用Neural Network Promoter Prediction、Promoter SCAN、Promoter 2.0和Meth Primer软件对启动子结构进行分析,PLACE在线分析预测可能的顺式作用元件。结果表明:克隆获得了一段长度为2 046 bp的Wcor15基因上游调控区序列,软件预测显示该区域含有4个可能的启动子,除了具有一般启动子的CAAT-box、TATA-box典型结构外,还含有CBFHV、ACGTATERD1、ABRELATERD1、ARR1AT、CANBNNAPA应答非生物胁迫、激素等重要作用元件。序列比对发现小麦Wcor15启动子与拟南芥cor15a、大麦cor14b的一致性分别为36.92%和40.29%。     

滇龙胆中细胞色素P450还原酶基因的克隆及生物信息学分析

细胞色素P450还原酶在体内可催化许多反应,通过FAD和FMN辅基将NAD(P)H的电子传递至细胞色素P450酶,从而使细胞色素P450酶同底物产生氧化还原反应,起到电子供体的作用.本研究对药用植物滇龙胆(Gentiana rigescen Franch.)的细胞色素还原酶基因GrCPR1进行克隆、生信分析和异源表达....     

地黄己糖激酶RgHXK2基因的电子克隆及生物信息学分析

己糖激酶(HXK)的主要功能是在糖酵解途径中催化己糖磷酸化,是植物体内调控呼吸代谢的关键酶之一。主要催化己糖磷酸化进入糖酵解途径,可在植物体的生长发育过程中为其提供大量的能量。己糖激酶属于hexokinase-2超家族,在植物生命活动中发挥着重要的作用。为了解地黄(Rehmannia glutinosa)己糖激酶基因RgHXK2在磷酸化过程中起到的作用,该研究基于转录组测序获得的地黄己糖激酶基因ORF的部分保守序列和地黄的SRA数据库,在NCBI中进行电子克隆获得地黄RgHXK2全长cDNA序列;采用在线网上工具ProtParam、TMHMM、SOPMA、SMART、MEME、Clustal Omega以及Jalview、MegaX等软件对其进行生物信息学分析,结果显示,地黄RgHXK2基因全长1 462 bp,包含1 194 bp的开放阅读框,编码一个含有397个氨基酸、相对分子质量为43.75 kD、等电点为6.34的蛋白质。在线软件预测该蛋白含有Hexokinase-1和Hexokinase-2保守结构域,无信号肽与跨膜结构,为亲水性非分泌蛋白,主要分布于线粒体和叶绿体中,参与碳...     

桑葚MaMYB44基因的克隆与生物信息学分析

本研究对桑葚(Morus alba L. cv.‘Cheongil’) MaMYB44转录因子进行了生物信息学等的研究,首先对桑葚中MaMYB44基因进行克隆。通过生物信息学分析,发现MaMYB44基因长度为720 bp,编码239个氨基酸,分子质量为58 215.61 Da,理论等电点5.10;通过分析桑葚蛋白MaMYB44的疏水性可知,其中的氨基酸呈现疏水性;通过桑葚MaMYB44蛋白进行跨膜区分析表明,Ma MYB44所有氨基酸都在细胞膜外,无跨膜结构,所以并不是跨膜蛋白;即使有信号肽结构的存在,但是螺旋结构并不存在;然后构建植物表达载体,获取阳性质粒,亚细胞定位后显示MaMYB44基因定位于细胞核中;进一步RT-qPCR分析得出结果,MaMYB44基因在茎中的表达量最高,其次是果实、叶、根。可初步推测该基因主要在桑葚茎中表达并行使功能。     

玉米ZmZFP36基因的克隆与生物信息学分析

玉米(Zea mays)是世界上最重要的粮食作物之一,研究其抗逆性机理有着十分重要的意义.本研究用在线软件预测了玉米miR408的靶基因,对其中编码锌指蛋白的靶基因ZmZFP36进行了生物信息学分析,并构建了该基因的植物表达载体.研究结果表明,ZmZFP36的转录本全长为1169 bp,无内含子,开放阅读框长度为702...     

小麦株高近等基因系的RAPD标记研究

采用ＲＡＰＤ技术,以４个小麦株市基因近等基因系（分别含有rht、Ｒht1、Rht2、Rht3)为材料,对２９６个单一随机引物（１０个核苷酸）进行了筛选。发现２５个引我的扩增产物的近等基因系间表现出特异性,在６次重复试验中,有１８个引物的特异护增片段不能重复,６个引物可以重复２－３资,唯有ＯＰＡＭ０１在全部试验中均能稳定重复,其特异扩增版段ＯＰＡＭ０１１８６０可以作为rht基因的ＲＡＰＤ标记。     

小麦供锌状况对叶片结构及叶绿体超微结构的影响

锌缺乏或过量使小麦叶肉细胞变小,多环复式细胞显著减少。缺锌植株的叶绿体中基粒垛数少,基粒垛叠片层少,基质片层少,部分片层膨胀;叶绿体内包含较大的液泡和淀粉“泡”;叶绿体被膜模糊,甚至消失.锌过量则使小麦的叶绿体变小,基粒和基质片层明显减少,部分片层膨胀,亲锇颗粒多;在叶绿体周围线粒体出现较多.     

Performance of Spelt (Triticum spelta L.) and Wheat (Triticum aestivum L.) at two Different Seeding Rates and Nitrogen Levels under Contrasting Environmental Conditions

The aim of this investigation was to obtain information about differences between spelt and wheat in relation to their performance under low-input conditions. Two spelt varieties ( Oberkulmer, Hercule ) and two wheat varieties ( Anna, Iend ) were grown at two different seeding rates (S1 = 200; S2 = 400 kernels/m²) and two nitrogen levels (Nl = 80 kg N/ha; N2 = 110 kg N/ha) at two contrasting locations (Muri; altitude 459 m asl; Oberwallestalden: altitude 1011 m asl). No growth regulators and fungicides were applied.
The overall mean for the "husked" yield (grains and glumes) of spelt was not significantly different from the grain yield of wheat. At the higher site Oberwallestalden however, the "husked" yield of spelt was 10.5 % higher than the grain yield of wheat. Although the number of ears per m² was higher at S2 than at S1, spelt and wheat produced a similar yield at both seeding rates (spelt: S1 = 96 % of S2; wheat: S1 = 95 % of S2) due to the higher single ear weight at S1.
The reaction of spelt and wheat to nitrogen was similar. The yield was 10 % higher at N2 than at Nl for both species. The performance of spelt under low-input conditions was better than for wheat in marginal areas for cereal production only.     

Nitrate Assimilation and Nitrogen Harvest in Uniculm 'Gigas' Wheat (Triticum aestivum L.)

Contribution of the upper laminae designated as 7, 8 and 9 (flag), to the total reduced nitrogen was determined by two different methods; integration of nitrate reductase activity over the entire growing season of individual laminae, and laminae removal at ear emergence. The values obtained by these methods were: 20, 14, and 12 %, and 23, 15, 12 %, respectively. The in vivo nitrate reductase activity was relatively higher in the lamina 9, which contributed an extra 8 % towards total reduced nitrogen when the other laminae were removed from the shoot. No such compensatory response was observed when either lamina 7 or 8 was retained.     

花药培养在小麦耐盐育种上的应用研究

在小麦花药培养中，采用加入一定浓度ＮａＣｌ（０．１％、０．３％、０．５％）的筛选培养基，用恒定浓度筛选法和逐渐增加浓度筛选法，可筛选出耐盐变异体，获得耐盐花培株系。两种筛选方法比较，逐渐增加浓度筛选法可以明显提高筛选效率。经５个有性世代的鉴定表明，耐盐变异体的耐盐特性可以稳定遗传给后代。经筛选的花培株系中，耐盐特性能够稳定遗传的株系比例约占１／４。在育种实践中，经多点试验鉴定，已选育出一批有实用价     

小麦株高近等基因系的RAPD标记研究

采用RAPD技术, 以4个小麦株高近等基因系(分别含有rht、 Rht1、 Rht2、 Rht3基因)为材料, 筛选了296个单一随机引物(10个核苷酸)。 发现25个引物的扩增产物在近等基因 系间表现出特异性。 在6次重复试验中, 有18个引物的特异扩增片段不能重复, 6个引物 可以重复2～3次, 唯有OPAM01在全部试验中均能稳定重复, 其特异扩增     

Die Ertragsbildung von Dinkel (Triticum spelta L.) und Weizen (Triticum aestivum L.) unter verschiedenen Umweltbedingungen im Freiland

Dry Matter Production of Spelt ( Triticum spelta L.) and Wheat ( Triticum aestivum L.) under Different Environmental Conditions in the Field
The primary aim of our work was to obtain information regarding the germination characteristics of spelt under different environmental conditions in field experiments, and to trace possible differences between spelt and wheat, with regard to crop production. For the present study two spelt varieties ( Ostro/Rouquin) and two wheat varieties ( Arina/Probus) were used. In addition the spelt cultivars Altgold, Oberkulmer and Hercule were included in some of the experiments. To obtain information on the influence of the glumes, spelt was planted in the hulled (SP +) and dehulled (SP-) form.
Field emergence of spelt SP- was up to 40 % lower than that of SP+, Under water logged soil, the percentage of germination was higher for spelt SP+ than for wheat.
Spelt produced more tillers per plant than wheat. However, tiller reduction was also higher for spelt so finally both species had similar numbers of ears per plant. Plants with a high tiller production might be more flexible in response to unfavourable growing conditions.
Grain yield of spelt and wheat was developed in different ways. Spelt produced fewer but larger kernels per ear, whereas wheat produced more but smaller kernels. In 1986 grain yield was up to 20 % higher in wheat than in spelt. In 1987, an unfavourable year for cereal crop production in marginal areas, grain yield was higher for spelt than for wheat. For the farmer, the "husked" form of spelt is economically important. This was equal or higher than the grain yield of wheat in both years.
The high germination rate under unfavourable conditions, in combination with a high tillering capacity and the production of large kernels appear to be the major reason for the stable yielding ability of spelt.     

普通小麦(Triticum aestivum L.)T型细胞质雄性不育系及其保持系的线粒体DNA比较研究(英文)

mtDNAs of T type wheat cytoplasmic male sterile lines Ning Drawf 14(ND14) and their maintainers Ning Drawf 13 (ND13) were isolated and digested completely with restriction endonucleases EcoRI, PstI, EcoRV, BamHI. The results revealed that the molecular structure of mtDNAs from ND14 and ND13 cytoplasms were significantly deviated. The mitochondrial genomic difference between CMS line and maintainers were uncovered by southern hybridization with probes of 18S 5S rRNA、atpA genes from wheat and pea mitochondria, respectively. Due to the complexity of mtDNA and no proof of protein difference, it has not yet been demonstrated whether mtDNA difference of Normal and Male Sterile Cytoplasm of wheat is associated with CMS.     

Agronomic and Physiological Study of Cold and Flooding Tolerance of Spelt (Triticum spelta L.) and Wheat (Triticum aestivum L.)

Flooding of soil occurs all over the world in areas of crop production. The resulting lack of available oxygen in the soil can hinder the establishment of field crops such as wheat ( Triticum aestivum L.) and thus lead to a decrease in yield. The current study compares the tolerance of spelt ( Triticum spelta L.) and wheat to wet and cold stress during germination and early growth. Two specific traits seem to be related to the superior early flooding tolerance of spelt: (1) fast growth of the coleoptile, which reaches the soil surface rapidly, thus reducing the time span of hypoxia sensitivity, independent of temperatures between 10 and 20 °C; (2) the improved physiological adaptation for coleoptile growth under hypoxia is probably due to lower oxygen consumption between germination and emergence. Because of this high level of flooding tolerance, spelt would also be a good source for breeding for stress tolerance in wheat. These findings will help to detect quantitative trait loci (QTL) for characters such as growth rate, flooding tolerance and oxygen consumption in future studies. A marker-assisted introgression of spelt into wheat is necessary to avoid combination with undesirable agronomic traits of spelt.