碱茅Put-HVA1基因的克隆及在酵母和拟南芥中的表达

本研究从碱茅(Puccinellia tenuiflora)cDNA文库中分离得到Put-HVA1基因,其开放读码框(ORF)长534bp,共编码177个氨基酸,其预测分子量约为18.167kD,理论等电点约为7.80。氨基酸序列比较结果表明,Put-HVA1氨基酸序列与水稻、玉米的HVA1蛋白氨基酸序列的同源性分别为69%和56%。另外,将Put-HVA1基因构建到酵母表达载体pYES2和植物双元表达载体pBI121上,并分别转化至酵母(INVSc1)和拟南芥(Arabidopsis thaliana)。对pYES2-Put-HVA1转化酵母进行盐碱、氧化胁迫、渗透胁迫及干旱处理,结果表明:重组酵母提高了对盐碱、氧化、渗透胁迫及干旱等逆境的适应能力。对部分转基因株系进行了抗盐性试验,表明转Put-HVA1基因拟南芥在150mmol/L NaCl和5mmol/L NaHCO3胁迫下长势优于野生型拟南芥,表现出一定的耐受性。     

大豆Gm TIP1;1基因的克隆与表达分析

利用RT-PCR方法从大豆根部组织获得Glyma03g34310.1开放阅读框(ORF)全长, 经测序验证、Blast比对与同源性分析发现该序列编码的蛋白质与其他植物的TIP1;1蛋白具有较高的相似性, 故命名为GmTIP1;1基因(GenBank登录号为AK285481), 该基因ORF长753 bp, 编码1个包含250个氨基酸的蛋白, 在ORF内部第381个核苷酸处含有1个94 bp的内含子, 符合↓GT--AG↓的剪接方式; 系统进化树分析发现Gm TIP1;1聚类到豆科植物分支, 其他不同科的植物也有规律地聚到了不同分支, 推测该蛋白氨基酸序列可以作为植物分类的依据; 半定量RT-PCR结果表明该基因在大豆的不同器官、不同器官的不同发育阶段均具较高且同等的表达水平, 暗示该基因在植物的整个发育进程中均具重要作用; 在盐胁迫的不同时间点其表达量有下降的趋势, 但仍然保持较高的表达水平; 以pYES2为酵母表达载体, 转化酿酒酵母INVSc1菌株, 获得重组酵母INVSc1 (pYES2-GmTIP1;1), 转化菌株在盐胁迫下的存活率明显高于对照INVSc1 (pYES2), 而在干旱胁迫下则没有显著差异, 表明该基因的表达能有效地提高酵母的耐盐性。     

紫杆柽柳cDNA文库构建与硫氧还蛋白基因的克隆

以NaHCO3胁迫的紫杆柽柳(Tamarixandrossowii)为材料建立了cDNA文库。经检测,文库的初始滴度为5.7×105pfu,重组率为96%,插入片段的长度在0.3~3.0kb之间,平均长度为1.2kb,扩增后文库的滴度为4.0×109pfu/ml。对文库克隆进行随机测序,获得了紫杆柽柳的硫氧还蛋白(Thioredoxin)基因序列。生物信息学分析表明获得的硫氧还蛋白基因全长683bp,其中5'非翻译区长56bp,3'非翻译区长204bp,开放读码框长423bp,编码140个氨基酸,蛋白的分子量为15.1kD,理论等电点为5.59,其氨基酸序列中具有硫氧还蛋白的保守活性位点“WCGPC”序列,Clustal分析表明该硫氧还蛋白与蓖麻(Ricinuscommunis)的硫氧还蛋白氨基酸序列同源性高,达65%。本研究构建的cDNA文库为柽柳基因克隆和系统研究柽柳抗逆分子机理奠定了基础。     

刚毛柽柳液泡膜H~+转运无机焦磷酸酶ThVP3的克隆与表达分析

通过对刚毛柽柳转录组的分析,获得并克隆了1个H~+-PPase基因cDNA全长,命名为ThVP3。序列分析结果表明:ThVP3全长3 500 bp,包含2 406 bp完整的开放读码框,618的5'非编码区和476 bp的3'非编码区。该cDNA编码1个801个氨基酸的多肽,等电点为5.67,推测分子质量为84.86 k D。通过与其他物种的氨基酸多序列比对及系统进化树分析,发现ThVP3氨基酸序列与AtVP2(拟南芥)一致性达88%,属于Ⅱ型VP成员。ThVP3基因有15个跨膜区,并具有高度保守的3个结构域(CS1,CS2,CS3)。相对荧光定量RT-PCR结果表明,ThVP3在高盐及干旱胁迫下均呈现为表达上调,但在各胁迫时间点和不同组织中的表达模式有所差异,0.4 mol/L Na Cl胁迫处理下,地下部分3 h达到最大值后有不同程度的降低,地上部分24 h达到最大值,与地下部分比,地上部分呈现较高的表达量;20%(w/v)PEG 6000胁迫处理下,地下部分在3 h和12 h达到峰值,地上部分表达量逐渐升高,24 h达到最大值后逐渐降低,96 h达到第2个峰值。ThVP3的表达受盐旱胁迫的诱导,推测其可能在柽柳的胁迫应答中起着重要的作用。     

棉花亲环素基因(GhCYP1)克隆及在干旱胁迫下的表达分析

本研究从已构建的棉花(Gossypium hirsutum)干旱胁迫抑制性差减杂交(suppressive subtractive hybridization,SSH)cDNA文库分离得到一个含有植物亲环素保守结构域的EST片段,测序并结合cDNA末端快速扩增(5'-RACE)技术获得了1个779bp的cDNA序列。序列分析表明,该cDNA5'非翻译区为70bp,3'非翻译区为187bp,并含有一个编码174个氨基酸蛋白的开放阅读框。Blast分析表明,该基因的编码产物为一个亲环素蛋白,将该基因命名为为GhCYP1,序列提交到GenBank,登录号为GQ292530。半定量RT-PCR分析表明,干旱胁迫处理后,该基因在叶片中的表达量迅速提高,并在胁迫2h达到最高,这一研究暗示该基因的表达与棉花抗旱胁迫相关。     

虹鳟鱼NDK-1 基因cDNA的克隆与序列特征分析

以虹鳟鱼(Oncorhynchus mykiss)为研究材料,使用RT-PCR和RACE法分离和克隆了虹鳟鱼NDK-1基因cDNA的全长序列(GenBank登录号:(FJ607868).序列全长953 bp,其中5'端非翻译区(5'-UTR)长45 bp,3'端非翻译区(3'-UTR)长284 bp,开放性阅读框(ORF)长624 bp,编码207个氨基酸.虹鳟鱼NDPK蛋白序列与几种脊椎动物台湾樱花钩吻鲑、斑马鱼、西方爪蟾、非洲爪蟾、人和黑青斑河的同源性分别为95%,72%,68%,68%,65% 和 64%,表明NDK-1基因在进化过程中是高度保守的.     

棉花转录因子基因GhMYB的克隆及特征分析

从棉花中克隆了1个MYB基因,命名为GhMYB(Gen Bank No.HQ234875)。该基因c DNA全长1264bp,具有1个774 bp的开放阅读框,5'非编码区为250 bp,3'非编码区为240 bp,编码256个氨基酸,预测分子量约为28.867 k Da,等电点为8.56,推测的氨基酸序列中含有2个高度保守的SANT结构域。GhMYB基因组序列长度为1355 bp,包含2个外显子和1个内含子。氨基酸同源分析发现,GhMYB与来自可可、陆地棉和巴旦木的MYB同源性较高。亚细胞定位结果表明,GhMYB:GFP融合蛋白定位于细胞核中。SDS-PAGE电泳分析表明,p ET-28a-GhMYB最佳诱导表达条件为1.0 mmol·L-1 IPTG在37℃下诱导4 h。电子表达谱分析表明,GhMYB基因在棉铃中特异表达,在根和叶中受水涝胁迫的诱导表达。     

TcLr35小麦β(1,3;1,4)葡聚糖苷酶基因cDNA全长的克隆及分析

根据已发表的植物β(1,3;1,4)葡聚糖苷酶基因设计引物,利用RT-PCR和RACE技术,从被小麦叶锈菌诱导的小麦抗叶锈病基因近等基因系材料TcLr35中获得一个β(1,3;1,4)葡聚糖苷酶基因cDNA全长,命名为PR34,该基因全长序列为1416bp,具有一个编码335个氨基酸的开放阅读框(ORF),其起始密码子ATG位于13bp处,终止密码子TAG位于1015bp处,3'末端有20bp的poly(A)尾,379bp的3'非翻译区。与GenBank中小麦、大麦等多个葡聚糖苷酶基因具有较高同源性;对其推断的氨基酸序列进行分析,含有Glyco_hydro_17保守结构域,为葡聚糖苷酶基因蛋白结构域。Southern杂交显示,该基因在TcLr35小麦基因组中为低拷贝。     

紫花苜蓿几丁质酶基因MsChiⅣ克隆及序列分析

本研究根据截形苜蓿(Medicago truncatula)根部的几丁质酶基因保守序列(GenBank登录号:AF167328)设计引物,通过RT-PCR直接扩增的方法得到了紫花苜蓿(Medicago Sativa L.)根部几丁质酶基因保守序列.根据得到的保守序列设计3'端和5'端特异引物,分别从3'端和5'端扩增延长该片段,最后通过序列拼接获得紫花苜蓿根部一种几丁质酶基因的全长cDNA序列,命名为MsChiⅣ(GenBank登录号:FJ487629).MsChiⅣ基因全长为1 025 bp,开放性阅读框全长为849 bp,编码282个氨基酸,分子量为30.5 kD,预测等电点为4.66,其结构包括信号肽、几丁质结合区、糖苷水解酶区,为Ⅳ类几丁质酶,属于几丁质酶第19家族,在氨基酸水平上与截形苜蓿几丁质酶蛋白同源性最高,达到98%.蛋白结构分析表明该基因编码蛋白为非跨膜蛋白,存在于细胞质中.     

小麦脂质转运蛋白cDNA结构及相应肽链构造特征的分析

根据测序获得的1条261 bp cDNA片段,通过电子延伸、设计引物,从小麦Mardler/7*百农3217(Pm2)的cDNA中扩增获得1条651 bp的cDNA片段,通过同源比对发现其包含小麦LTP1 完整的编码序列.该片段包含基因的5'非翻译区42 bp,3'非翻译区261 bp,开放阅读框348 bp,编码115个氨基酸.预计蛋白的分子量为11.2 kD,等电点为9.46.此基因有8个位置保守的半胱氨酸(C)残基及25个氨基酸的信号肽,为典型的植物脂质转运蛋白基因.其基因序列数据库(GenBank)登录号为AY796184(基因)和AAV65513(蛋白).通过疏/亲水性分析,发现肽链分子具有较大范围的疏水面.     

Location of a high-lysine gene and the DDT-resistance gene on barley chromosome 7

Summary The high-lysine gene in Risø mutant 1508 conditions an increased lysine content in the endosperm via a changed protein composition, a decreased seed size, and several other characters of the seed. The designation lys3a, lys3b, and lys3c, is proposed for the allelic high-lysine genes in three Risø mutants, nos 1508, 18, and 19. Linkage studies with translocations locate the lys3 locus in the centromere region of chromosome 7. A linkage study involving the loci lys3 and ddt (resistance to DDT) together with the marker loci fs (fragile stem), s (short rachilla hairs), and r (smooth awn) show that the order of the five loci on chromosome 7 from the long to the short chromosome arm is r, s, fs, lys3, ddt. The distance from locus r to locus ddt is about 100 centimorgans.     

Simultaneous Determination of Four Phenolic Acids in Radix Salviae Miltiorrhizae Formula Granules by QAMS

[Objectives]This study aimed to establish a QAMS(quantitative analysis of multi-components by single-marker)method for simultaneous determination of four phenol...     

Pollen and pollination experiments. III. The viability of apple and pear pollen as affected by irradiation and storage

Summary Apple and pear pollen was irradiated with doses of 0, 50, 100, 250 and 500 krad (gamma rays) and stored at 4°C and 0–10% r.h. From the in-vitro germination percentages an average LD 50 dose of about 220 krad was estimated. For both irradiated and untreated pollen a close and corresponding lineair relationship existed between germination percentage and pollen tube growth.Irradiated pollen was much more sensitive to dry storage conditions than untreated pollen, resulting in less germination and more bursting. Apparently, irradiation caused the pollen cell membrane to lose its flexibility faster than normal. Rehydration of dry-stored, irradiated pollen in water-saturated air restored germination percentages up to their initial levels. The importance of this procedure in germination trials is stressed.     

Evolution under domestication involving disturbance of genic balance

Summary An hypothesis is developed that the rapid change from wild plants into domesticated crops principally involves the selection of alleles with non-functional gene products which leads to reduced control of the highly integrated metabolism and morphogenesis previously accumulated by lengthy natural selection. Such disturbance of the genome produces altered physiological and morphological development which, although deleterious in nature, serves mankind better and has been selected.     

Genetic variation for disease and nematode resistances and forage quality in perennial diploid and tetraploid lucerne populations (Medicago sativa L.)

Summary Twenty-five lucerne populations of the Medicago sativa complex, which were either diploid or tetraploid and wild or cultivated, were analysed for their resistance to four different fungal diseases and to stem nematode. Forage quality, including stem digestibility and saponin content, was also tested.Populations varied in susceptibility to the diseases caused by Colletotrichum trifolii, Verticillium albo-atrum, Sclerotinia trifoliorum and Pseudopezizza medicaginis, and to the nematode Ditylenchus dipsaci. Except for Sclerotinia rot, sativa and falcata subspecies differed in susceptibility, but this grouping of populations did not account for the full range of variation among them. However, the resistance to P. medicaginis was much lower in the sativa than in the falcata populations.Populations also varied significantly in stem fiber content and digestibility. Stem digestibility was negatively correlated to forage yield. Wild sativa and falcata populations had lower fiber content and higher digestibility than cultivated sativa populations. The medicagenic acid was the sapogenin responsible for the anti-nutritional effect of the lucerne measured by the yellow mealworm larvae Tenebrio molitor. The medicagenic acid content was lowest for the pure sativa populations, highest for the pure falcata populations, and intermediate for the French sativa varieties that have some traits originating from falcata germplasm. Some populations could be used in breeding programs to improve disease and nematode resistance, and forage quality.     

Review of Effects of Active Components of Gastrodiae Rhizoma on Central Nervous System

In order to explore the pharmacological effects of active components of Gastrodiae Rhizoma on the central nervous system,through consulting related literatures,...     

Effects of Pb on Rhizosphere Soil Enzyme Activity and Chemical Constituents of Achyranthes bidentata Blume

[Objectives]The purpose of this study was to investigate the effects of Pb on rhizosphere soil enzyme activity and chemical constituents of Achyranthes bidentat...     

不同P-Zn配比对小麦幼苗微量元素营养的影响

采用了螯合-缓冲营养液培养方法对小麦进行了苗期培养试验,在3个P水平(0,0.6,3.0 mmol/L)和3个Zn水平(0,3,30 μmol/L)的完全组合下对小麦苗期生长及Zn、Fe、Cu、Mn营养进行了研究,旨在为小麦微肥施用提供理论依据.结果表明,P、Zn的正常供应促进了小麦生长,二者的缺乏与过量均会抑制小麦发育,且这种影响在冠部表现得更为明显.在小麦苗期,Zn与Cu的吸收存在明显的拮抗作用,但供Zn则促进了Zn和Cu的转运,而Mn转运则受到了抑制;过量供Zn时,大量Zn被转运到冠部,同时明显抑制了(Fe+Cu+Mn)的吸收总量;P的供应显著地抑制了Fe的吸收,但P的供应提高了Zn、Cu、Mn的转运率;P、Zn在对Zn与Fe、Cu、Mn间吸收竞争的影响中,Zn本身的影响要比P的影响更为明显,供Zn明显促进了小麦幼苗对Zn的吸收;在小麦幼苗冠部,Zn与Fe的竞争中,供P利于Zn的吸收,缺P则利于Fe的吸收;而Zn与Cu以及Zn与Mn间的竞争中,缺磷时利于Zn的吸收,供磷后则利于Cu和Mn的吸收.总之,小麦幼苗Zn、Fe、Cu、Mn营养中,P、Zn的不同配比会不同程度地改变Zn与Fe、Cu、Mn的协同或拮抗效应.     

Isozyme variation between diploid and tetraploid cytotypes of Glycine tabacina (Labill.) Benth.

Summary Glycine tabacina (Labill.) Benth. is a wild perennial species related to the cultivated soybean, G. max (L.) Merr. It is composed of diploid (2n=40) and tetraploid (2n=80) cytotypes. Currently, to differentiate the cytotypes, plants are grown out in the greenhouse and chromosome counts made on pollen mother cells. It is a laborious and time consuming process. The objective of this study was to determine whether electrophoretic techniques could be utilized to separate the cytotypes. Electrophoretic examination of seven isozyme systems from seed of 67 G. tabacina accessions revealed banding patterns that could be used to differentiate between diploid and tetraploid cytotypes in the species. Among the tetraploid accessions, the number of bands observed were always greater than the diploids. Some tetraploid banding patterns consisted of bands similar to the diploid tabacina and/or additional bands previously identified in other Glycine species. The patterns of isozyme multiplicity and variation in the tetraploid tabacinas suggests more than one mode of origin for the tetraploids.     

Some physiological and photosynthetic characters of rice × sorghum and rice × wheat hybrids

Summary Some photosynthetic characters as-leaf anatomy, leaf photosynthetic rate and CO₂ compensation pointdistinguishing C₃ and C₄ plants and physiological characters as leaf area ratio, nitrogen content, leaf stem ratio and total shoot-to-root and deep root-to-shoot ratios have been studied in rice × sorghum and rice × wheat hybrids. Rice × sorghum 1. has lower values of photosynthetic rate, leaf nitrogen, total root and deep root-to-shoot ratio and CO₂ compensation point as of rice parent where as, rice × sorghym 2. is superior in all these characters. Both hybrids lack kranz anatomy. Though both rice × sorghum hybrids show characters of C₃ rice plant but rice × sorghum 2. has improved drought tolerance and leaf characters in relation to yield. Rice × wheat hybrid have higher assimilatory area and higher total root-to-shoot ratio. Grains of rice × wheat hybrids are identical to rice grain. However, as grains of rice × wheat hybrid does not contain seed coat, it could be exploited as novel rice germplasm after improvement.Abbreviations T CO₂ compensation point - T21 CO₂ compensation point at 21% O₂ - T2 CO₂ compensation point at 2% O₂ - dR/sR deep Root-to-shoot Ratio - LAR Leaf Area Ratio - CER photosynthetic rate - TR/SR Total Root-to-Shoot Ratio