小麦SnRK2家族基因鉴定及进化分析

SnRK2是一种植物特异性的丝氨酸/苏氨酸蛋白激酶,在植物生长发育和胁迫耐受信号传递过程中发挥着至关重要的作用。为探索SnRK2在小麦抗逆中的作用,基于最新的小麦基因组信息,采用blastp和hmmer两种方法在普通小麦中共鉴定到30个SnRK2基因家族成员。系统发育分析表明,SnRK2基因在普通小麦及其祖先物种的进化过程中高度保守。利用RNA seq数据分析小麦SnRK2基因在不同组织以及不同胁迫条件下的表达模式,结果表明,小麦SnRK2基因家族成员在不同组织以及不同胁迫条件下表达量均存在差异,且具有显著的组织特异性。通过qRT PCR进一步验证6个小麦SnRK2基因的表达模式,发现不同基因之间的表达量存在明显差异,推测小麦SnRK2基因家族的部分成员出现功能分化。随后,利用已发表的六倍体小麦重测序数据,分析不同小麦群体中SnRK2基因的核苷酸多样性(P_i)和分化指数(F_st),并解析单倍型,推测Ta 5B SnRK2.28基因的AA单倍型为优异等位变异。通过同源建模预测小麦SnRK2蛋白的三维结构,发现小麦SnRK2蛋白结构在进化上高度保守。     

植物多功能调控因子SnRK2研究进展

蔗糖非发酵相关蛋白激酶2(sucrose non-fermenting 1-related protein kinase 2,SnRK2)是植物特有的一类丝氨酸/苏氨酸蛋白激酶,其主要通过磷酸化底物来调节下游基因表达及蛋白质活性。本文综述了蛋白激酶SnRK2的研究进展,包括其结构、分类、调控机制和生物学功能。SnRK2作为典型的多功能调节因子,不仅参与响应各种逆境胁迫,还参与调控植物生长发育。目前有关SnRK2的研究主要集中在ABA依赖信号通路,而对其在非ABA依赖信号通路中的研究很少,未来研究应该聚焦这一领域。SnRK2作用机理的全面解析对提高作物的抗逆性和改良产量性状都具有重要意义。     

蓖麻miR396基因家族及其靶基因GRF生物信息学分析及鉴定

MiR396-GRF在调控植物的生长发育和生物胁迫及非生物胁迫过程中发挥着重要作用,目前关于蓖麻miR396和GRF的研究还很少.本研究中利用BLASTP和Pfam在蓖麻基因组中鉴定到4个miR396和11个GRF成员.分析发现,RcGRFs转录因子蛋白长度在318-619aa之间,理论等电点在5.54-9.34之间,...     

番茄SlSnRK2s基因表达在促进叶片衰老中的作用

SnRK2(sucrose non-fermenting 1一related protein kinase2)是脱落酸(ABA)信号转导途径中的正调控因子,广泛参与了植物的生长发育、病虫害防御、ABA和非生物胁迫等多种信号的应答反应。从番茄中分离了3个与SnRK2.2、SnRK2.3和SnRK2.6同源的基因,分别命名为SlSnRK2.2、SlSnRK2.3和SlSnRK2.6,同时构建了SlSlSnRK2.2/2.3/2.6 RNAi载体,并通过根癌农杆菌介导的叶盘法转化番茄,得到转基因番茄植株。初步表型分析结果发现转基因植株呈叶片早衰的现象。通过对乙烯信号相关基因的检测可知,转基因植株中除了ERF2表达量低于野生型,其余3个基因(ACO4、ERF1B、ERF1)的表达量均高于野生型,表明SlSnRK2.2、SlSnRK2.3和SlSnRK2.6与叶片衰老有关,可能参与ABA和乙烯信号互作,此结果对揭示植物衰老机理具有重要意义。     

油棕C2H2基因家族鉴定及低温胁迫下的表达分析

C2H2型锌指蛋白是最常见的锌指蛋白之一,广泛存在于真核生物中,在植物的生长发育、盐、低温、干旱胁迫等非生物胁迫响应中扮演重要作用。本研究以拟南芥C2H2氨基酸序列作为探针,在油棕基因组数据库中进行本地BLASTP比对和保守结构域预测分析,筛选出油棕C2H2型锌指蛋白家族成员,利用生物信息学手段对基因家族成员的理化性质、进化关系、染色体位置、基因结构、保守基序和启动子顺式元件进行分析,并利用转录组数据和荧光定量PCR方法分析EgC2H2基因家族成员在低温胁迫下的表达情况。结果显示：从油棕基因组中共鉴定出73个C2H2型锌指蛋白家族成员,EgC2H2家族成员氨基酸的数目为128～556个,分子量为14176.73～58983.30Da,等电点为4.99～9.62,不稳定系数为38.84～86.01,亲水性为–1.154～-0.226,亚细胞定位显示C2H2基因家族所有成员均定位于细胞核;系统进化树分析显示EgC2H2家族成员可以分为4个亚家族,EgC2H2家族成员多数归类于第Ⅰ亚家族和第Ⅱ亚家族,且同一亚家族成员的结构域具有较高的一致性;染色体定位结果显示,EgC2H2家族成员不均匀地分...     

辣椒几丁质酶类基因家族的全基因组鉴定和表达特征分析

几丁质酶（chitinase, EC 3.2.1.14）是一种降解几丁质的糖苷酶,在植物应对非生物和生物胁迫中起重要作用。本研究对辣椒几丁质酶类（Capsicum annuum chitinase-like, CaCTL）基因家族成员进行了全基因组注释、进化分析和基因表达模式分析。从最新的辣椒基因组数据库中鉴定出31个CaCTL成员。根据系统发育进化树将这些成员分为GH18和GH19亚家族,并进一步分为5个亚类（Ⅰ~Ⅴ类）。保守基序分析结果表明,每个亚类中的成员存在功能相关性。对辣椒、矮牵牛以及番茄的CTL基因家族成员同源性分析表明,在矮牵牛和番茄中分别有7个和11个辣椒CaCTL的同源基因。通过qRT-PCR分析发现,在正常环境生长的辣椒植株中,64.2%的GH18亚家族的CaCTL成员表达水平很低,有64.7%的GH19亚家族成员在不同组织中存在差异表达。顺式作用元件分析表明,CaCTL成员启动子区域存在许多激素反应以及生物和非生物应激相关元件。当植物遭受不同的非生物胁迫时,qRT-PCR结果显示,多数CaCTL成员表达上调。当植物遭受高温干旱条件时,多个CaCTL成员的表达上调明显。上述结果丰富了CTL基因家族进化的研究,为探索CaCTL成员的功能提供了数据基础。     

蓖麻花序特征及花芽分化的初步研究

本文对蓖麻花序特征、顶芽分化及内源IAA和ABA含量变化进行初步研究,结果表明：不同品种（系）间雌花率及雌花密度均存在极显著差异。蓖麻雌花并非完全单性花,各品种（系）雌花均存在退化雄蕊痕迹,但其发生率不同,单雌后代品系明显高于品种材料。单雌花序与两性花序的雌花中均存在退化雄蕊,且无显著差异,但花序中下部与上部之间差异显著。顶芽石蜡切片观察表明：蓖麻在5叶期以前,其顶芽基本处于叶芽生长阶段;在6-9叶期,顶芽逐渐进入花芽分化、生长阶段,而此阶段,顶芽内源IAA、ABA含量变化也十分明显。不同性别花序和花的内源激素含量测定结果显示：IAA含量,单雌花序＞雌花＞两性花序＞雄花;ABA含量,雄花显著高于雌花,但两性花序却明显低于单雌花序;IAA/ABA值,雌花＞单雌花序＞两性花序＞雄花;说明IAA含量和IAA/ABA相对含量可能在花性分化中具有较为重要的作用。     

蓖麻种芽腐烂病害的病原鉴定

为明确引起蓖麻种芽腐烂病害的病原菌种类,本研究采用常规组织分离法分离获得病原菌,根据柯赫氏法则对病原菌进行致病性测定,并结合形态学特征观察和分子生物学技术确定病原菌分类。结果表明,从蓖麻种芽病样组织中分离得到RFR1-4菌株为致病菌。形态学鉴定该菌为茄腐镰孢菌(Fusarium solani)。对致病菌株进行ITS、β-tubulin和TEF-1α基因分析,结果显示与Fusarium-ID数据库中的茄腐镰孢复合种(F. solani species complex)有100%的相似性。因此结合形态学鉴定结果,确定引起蓖麻种芽腐烂病害的病原菌为茄腐镰孢复合种。该病原鉴定对蓖麻种芽腐烂病害的防控有重要的指导作用。     

蓖麻组织培养和植株再生的研究

本研究建立了一种有效的蓖麻植株再生体系。以蓖麻成熟种子胚轴为外植体,在含6-BA（0.5mg/L）的MS培养基上每个外植体均能诱导出由若干不定芽组成的丛生芽,平均每个外植体可产生6.3个不定芽,健壮的不定芽经伸长、生根而发育成完整的再生植株并能全部移栽成活。本实验所用的3个蓖麻品种在产生不定芽的能力和数量上没有明显差异。利用该体系所诱导的不定芽具有生长健壮和易于生长发育等特点。该再生体系的建立为蓖麻遗传转化研究奠定了基础。     

木薯MePP2CAb基因克隆及表达分析

2C型蛋白磷酸酶（protein phosphatase 2C,PP2C）在ABA核心信号途径中发挥着重要作用。为了研究PP2C基因在木薯响应非生物胁迫过程中的功能,本研究通过RT-PCR技术,从木薯Arg7中克隆得到MePP2CAb基因,并对其进行生物信息学分析、自激活活性分析、启动子活性分析及不同逆境和激素处理下的表达模式分析。结果表明：（1）MePP2CAb基因的长度为1296 bp,包含431个氨基酸残基,蛋白相对分子量和理论等电点分别为47.08 kDa和5.5,具有PP2C家族的结构域特征。蛋白质序列比对结果显示,MePP2CAb与橡胶树和麻风树的PP2C序列最为相似,一致性分别为82.75%和74.01%,在C-端保守。上述结果证明MePP2CAb基因属于PP2C家族。（2）MePP2CAb基因在木薯块根中的表达最高。（3）MePP2CAb具有一定的自激活活性,MePP2CAb基因的全长启动子的活性也较高。（4）MePP2CAb基因属于ABA核心通路,启动子序列分析显示,它包含ABRE(abscisic acid responsiveness)元件、MeJA响应原件、干旱...     

蓖麻三磷酸甘油脱氢酶基因 (RcGPDH)的克隆及功能分析

为了阐明蓖麻三磷酸甘油脱氢酶基因（glycerol-3-phosphate dehydrogenase, RcGPDH）在蓖麻油累积过程中的作用,本研究根据已报道的蓖麻种子表达序列标签（Expressed Sequence Tags,EST）设计引物,通过RACE（rapid-amplification of cDNA ends)方法克隆蓖麻RcGPDH基因的全长并对其序列进行分析,构建了该基因的酵母表达载体PYES2.1/V5-His-TOPO-RcGPDH,以载体PYES2.1/V5-His/lacZ质粒DNA为对照,转化酵母野生型菌株BY4742,运用分光光度法测定转基因酵母的生长曲线,通过香草醛法测定稳定生长期的转基因酵母的油脂含量。结果表明,转基因菌株比转空载体对照菌株生长慢,两者均在培养18h后进入平台期;两个菌株的油脂含量没有明显的差别,表明RcGPDH基因对酵母油脂的累积没有起到积极的作用,在蓖麻种子中可能还存在另一个GPDH同源基因参与TAG的合成。     

The feeding value of roasted castor oil bean (Ricinus communis) to growing chicks

A nutritive evaluation of roasted castor oil bean (Ricinus communis) was made using 150 day old crossbred progeny of barred Plymouth rock × Nigerian local chicken. The chicks were divided into five groups of 30 birds each. Each group was fed one of five iso-nitrogenous and iso-caloric diets A, B, C, D and E containing 0% (control), 10% non-roasted castor oil bean seeds (NRCOB), or 10, 15 or 20% roasted castor oil bean seeds (RCOB), respectively. Roasting was done at 140°C for 20 minutes in order to destroy the ricin component of the castor oil bean seed. There were significant (P<0.05) differences in average feed intake, daily gain, and feed conversion ratio between birds among the treatments. The best results were obtained with inclusion of RCOB at the 10% level. The birds fed the 20% roasted bean diets had the worst performance. The diet containing the 10% NRCOB supported little or no growth, and also resulted in a high mortality (83%) among the birds. There were no significant (P<0.05) differences in mortality rates between the control diet and the other diets containing RCOB.     

蓖麻DNA糖基化酶的序列与基因表达分析

为揭示DNA糖基化酶在调节蓖麻生长发育和影响基因印记过程中的分子机理,本研究基于蓖麻全基因组、利用生物信息学的方法结合转录组数据和RT-PCR半定量表达分析,鉴别蓖麻DNA糖基化酶氨基酸序列的结构特征、系统发生和表达形式。结果显示：三种（DME、ROS和DML）DNA糖基化酶氨基酸均为不稳定氨基酸,具有典型的与DNA结合和碱基切除修复有关的保守结构域;系统进化分析发现三种DNA糖基化酶在植物中同源性强,而且是独立演化的结果;在不同组织的表达上,蓖麻的DME和ROS基因具有相似的表达形式,即在根、茎和叶中不表达,在胚乳中表达最高,而DML在各个组织中都未检测到基因的表达。     

Leaf gas exchange, chloroplastic pigments and dry matter accumulation in castor bean (Ricinus communis L) seedlings subjected to salt stress conditions

Brazilian castor bean (Ricinus communis L) crops have expanded towards semiarid lands, in which soil salinity is an important factor limiting plant development. In order to evaluate the effects of salt stress on leaf gas exchange, pigments, and dry matter accumulation of seedlings, seeds of castor bean var. BRS Nordestina were planted in 15-L pots filled with 13 kg of soil (control) or with soil plus 2 g NaCl kg⁻¹ soil corresponding to 30 mM NaCl (salt treatment). Seedlings were grown under greenhouse conditions and the effects of salinity were assessed by measuring physiological parameters at 38 and 59 days after germination. Salt treatment induced decreases in leaf water potential at pre-dawn (ψ_pd, 42%), stomatal conductance to water vapor (g_s, 36%), and net carbon assimilation rate (A, 24%) only at 38 days after germination. At the same time, the values for transpiration rate were unchanged and the hydraulic conductance was increased (34%). After 59 days under hypersalinity, leaf ψ_pd increased to −0.44 MPa, although g_s, A, transpiration rate, and hydraulic conductance were not different between treatments. Salt stress effects on chlorophyll a, chlorophyll b and chlorophyll a + b contents were evident only on experimental day 59. The dry matter accumulation of leaves, roots and stems, as well as the total dry matter, and the root to above ground ratio increased with plant growth in both treatments, however, these parameters were lowered in salt-stressed seedlings. In short, all physiological variables were more drastically affected after 38 days under high salinity, and despite acclimatization of A after 59 days exposure to salt, no recovery of the dry matter accumulation was observed in the seedlings, suggesting that this species does not tolerate salt stress at its initial growth stages.     

蓖麻RcWD40家族鉴定与表达分析

植物中的WD40蛋白家族通过蛋白质-蛋白质及蛋白质-DNA的互作,参与生物过程,该家族的重要成员TTG1在拟南芥中抑制种子脂肪酸的过早积累。为揭示油料作物蓖麻中RcWD40基因家族及RcTTG1基因的表达特征,利用生物信息学手段在全基因组水平鉴定到182个RcWD40家族成员,其中RcWD40-181为RcTTG1基因,并对它们的系统发育、保守结构域及表达模式等进行分析。根据进化树结构和保守域组成分别将RcWD40家族成员分为8个cluster和28个亚家族。此外,转录组分析显示RcWD40在蓖麻的花、叶、根、茎以及各发育时期种子中具有多样的表达模式,预测家族基因可能调节这些组织的生长发育。启动子区域转录因子结合位点及种子发育表达分析结果预测RcTTG1可能承担与AtTTG1相似的脂肪酸积累负调节功能。     

Influence of spices on protein utilisation of winged bean (Prophocarpus tetragonolobus) and horsegram (Dolichos biflorus)

The influence of a mixture of eleven spices commonly consumed in India on the utilisation of protein from boiled winged bean (Psophocarpus tetragonolobus) and horsegram (Dolichos biflorus) was studied at 10 and 20 percent level of protein intake in experimental rats. Spices used in the mixture include red chillies (Capsicum annum), black pepper (Piper nigrum), coriander (Coriandrum sativum), cumin (Cuminum cyminum), garlic (Allium sativum), ajowan (Carum copticum), turmeric (Curcuma longa), caraway seeds (Carum carui) and fennel seeds (Foeniculum vulgare). Addition of this spice mixture at 1.5% level of the diet decreased the TD of both legumes, significantly only in the case of horsegram. A significant increase was observed in the BV of both the legumes at both levels of protein tested.     

Treatments for reducing total vicine in Egyptian faba bean (Giza 2 variety)

The response of faba bean Vicia faba (Giza 2 variety) towards soaking conditions differed greatly since the absorbed quantities of water (either by the whole or the decorticated forms) are a function of their chemical constituents. On the other hand, 28.45% of the total vicine (vicine + convicine) present in the whole faba bean samples was extracted after soaking for 72 h at room temperature. Subsequently, other soaking mediums, i.e., 0.5% sodium carbonate and/or 1% acetic acid were used in an attempt to increase the level of vicine elimination. Percentage removal of total vicine in whole faba bean was higher in the acidic (61.31%) than the alkaline (38.40%) medium under the conditions tested, i.e., at room temperature for 72 hours. The rates of vicine + convicine elimination in decorticated faba bean for the acidic acid and alkaline soaking media were 78.46 and 79.13%, respectively. The solubility ratio of total vicine relative to soaking solutions (H₂O:Na₂CO₃:Acetic acid) was 1:1.35:2.16 in the whole broad bean and 1:2.41:2.39 in the decorticated samples. The residual amounts of total vicine (78.33% and 77.27%) present after stewing under normal and under pressure cooking conditions could be expected to be decreased to 30.33% for the former and 29.92% for the later after 72 h of soaking. Regression analysis was used to estimate the theoretical zero point of vicine elimination from faba bean through soaking in 1% acetic acid.     

Comparative study of enzymes related to proline metabolism in tepary bean (Phaseolus acutifolius) and common bean (Phaseolus vulgaris) under drought and irrigated conditions,and various urea concentrations

There are several mechanisms used by plants for survival in adverse environments such as drought, high temperature and salinity. The objective of this study was to evaluate the drought tolerance of tepary bean as a function of biochemical processes linked to isozyme synthesis and changes in enzymatic activity related to proline metabolism. Mature seeds of common beans var. flor de mayo, Phaseolus vulgaris and tepary beans Phaseolus acutifolius were grown under two water conditions (irrigation and drought), and four levels of urea. Vertical electrophoresis and spectrophotometric techniques were used to evaluate protein patterns, glutamate dehydrogenase (GDH), proline oxidase (PO) and pyrroline-5-carboxylate reductase (P5C reductase) enzyme activities. These enzymes were studied because they are directly related to protein synthesis. Electrophoretic patterns showed more proteins in tepary beans than in common beans with limited irrigation. GDH showed only one isozyme, with a molecular weight between 240 to 270 kDa. A decrease in PO activity was observed in common beans under drought stress with a value of 237 mol/min, in comparison to irrigation conditions of 580 mol/min. GDH and P5C reductase enzymes have had higher activity in common beans than in tepary beans under water stress. There was a significant difference only in glutamate dehydrogenase enzyme with respect to urea level. The results suggest that drought tolerance of tepary beans is due to biochemical processes related to proline metabolic enzymes.