牡丹开花相关基因PsAP1的克隆与表达

APETALA1基因对花器官的形成具有重要作用,并且能够调节花期.以牡丹品种赵粉(Paeoniasuffru-ticosaL.cv.Zhaofen)为试材,采用RT-PCR和RACE方法从花瓣中获得了1个牡丹APETALA1基因cDNA全长,命名为PsAP1,GenBank登录号为HM143943.其cDNA全长1103 bp,包含130 bp的5′非编码区、244 bp的3′非编码区和1个长度为729 bp编码242个氨基酸的开放阅读框.序列比对和系统进化分析表明,PsAP1与葡萄的亲缘关系最近,相似性达80%以上,属于MADS家族AP1/SQUA亚家族.相对荧光定量PCR分析表明,PsAP1在花瓣中的表达量最高,在雄蕊中表达量最低.     

牡丹二氢黄酮醇4-还原酶基因PsDFR1的克隆及表达分析

二氢黄酮醇4-还原酶（DFR）是花色素苷合成途径中的关键酶,在植物花色的形成过程中起重要作用。本研究以牡丹品种‘彩绘’为试材,采用RT-PCR和RACE方法从花瓣中获得了一个牡丹DFR基因cDNA全长,命名为PsDFR1,GenBank登录号为HQ283448。序列分析结果表明,PsDFR1全长1242bp,包含一个长为1095bp的开放阅读框,编码364个氨基酸。生物信息学分析显示该基因编码的蛋白具有典型的DFR蛋白功能结构域,包括多个保守的短链脱氢/还原酶家族的特征位点,属于NADB_Rossmann超家族。氨基酸序列比对和系统进化树分析表明,PsDFR1与葡萄、棉花、毛果杨、梨等植物的DFR相似性都在75%以上。实时荧光定量PCR分析表明,PsDFR1在花色素大量积累的花瓣中表达量最高,其次是萼片和雄蕊,再次是叶片,在心皮中表达量最低。     

簸箕柳AP3同源基因SsMADS的克隆与特性分析

用cDNA末端快速扩增(rapid-amplification of cDNA ends,RACE)方法从簸箕柳雄花序中克隆了一个AP3同源基因的cDNA,长826 bp,包括完整的编码区、5′-UTR和3′-UTR,并将其相应的基因命名为SsMADS。该基因由7个外显子和6个内含子组成,编码区长723 bp,编码241个氨基酸,其N-端具有典型的MADS保守结构域。序列分析表明,SsMADS编码的氨基酸序列与毛果杨(Populus trichocarp)AP3同源蛋白有95.7%相似性,与其他几种柳属植物的AP3同源蛋白相似性达96.1%～99.6%。实时定量RT-PCR表明,SsMADS在叶、茎和根中表达量极低,在花序中表达量较高,并且其表达量在花器官的早中期发育阶段逐步提高,说明该基因在簸箕柳花器官的发育中起作用。     

羊草OEE1基因的克隆及盐胁迫下的表达

从羊草(Leymus chinensis )叶片cDNA文库中克隆得到可能编码33 kD的光系统Ⅱ(PSⅡ)外周蛋白(oxygen-evolving enhancer protein1,OEE1)全长cDNA(GenBank登录号为EF583851),命名为LcOEE1.序列分析结果表明,该cDNA全长1 107 bp,5′非编码区为32 bp,3′非编码区为71 bp,编码区长987 bp,编码328个氨基酸.BALSTp比对发现,该基因氨基酸序列与已报道的小麦和水稻中的OEE1序列具有95%和94%的相似性.聚类分析表明,该基因与小麦和水稻的亲缘关系较近,与拟南芥和菠菜OEE1基因的亲缘关系较远.Northern杂交结果表明,在200 mmol/L的NaCl处理7 d的幼叶中,OEE1 mRNA的表达量明显高于未处理的对照,说明羊草中OEEl基因受盐诱导.     

香蕉MaCAM基因克隆及表达分析

从香蕉根的cDNA文库中获得了一段香蕉钙调蛋白基因的片段,采用RACE技术获得其全长,命名为MaCAM。该基因全长845bp,编码149个氨基酸。生物信息学分析表明,该蛋白属稳定蛋白,其等电点为4.12,有2个保守的EFh功能结构域。与已知植物的钙调蛋白基因相比,一致性达90%以上。其中与粳稻、油棕、胡萝卜、甘蔗的CAM编码的氨基酸序列的一致性分别为99.33%、96.71%、98.00%、98.66%。系统进化树比对分析显示,香蕉与甘蔗的亲缘关系最为密切。器官特异性分析表明,MaCAM在香蕉的根、球茎、叶片、花和果实中均有所表达,在根中表达量最高,花中次之,而在叶片中的表达量最低。     

西伯利亚蓼钙调蛋白基因的克隆及其在盐胁迫下的表达

已从西伯利亚蓼叶中cDNA文库中获得的钙调蛋白EST序列,采用cDNA末端快速扩增（RACE）技术克隆了具有完整编码区的钙调蛋白基因的cDNA序列（GenBank登录号GQ988382）,命名为PsCaM。该基因全长615bp,编码区为450bp,编码149个氨基酸,5＇非翻译区为63bp,3＇非翻译区为102bp。同源性分析表明,该蛋白与其他植物钙调蛋白高度保守,氨基酸同源性高达98%。用实时荧光定量PCR研究3%NaHCO3胁迫下西伯利亚蓼基因表达的结果显示,自然条件下,该基因在叶中表达量最高,地下茎次之,茎中最低;盐胁迫下CaM在西伯利亚蓼的地下茎、茎和叶中均有表达,表达模式不同。     

苦荞查尔酮合酶基因CHS的结构及花期不同组织表达量分析

采用同源克隆、染色体步移和RT-PCR技术,首次克隆到苦荞查尔酮合酶基因(CHS)的全长DNA序列和cDNA开放阅读框(ORF)序列.序列分析表明,苦荞CHS DNA序列(GU172165)全长1 632 bp,含1个445 bp的内含子;cDNA编码区(HM852753)全长1 188 bp,编码395个氨基酸,命名为FtCHS.生物信息学分析表明,FtCHS和推导的氨基酸序列与其它植物CHS基因同源率在95%以上,含有CHS多基因家族的标签序列(GFGPG)、活性位点、底物结合口袋位点和环化反应口袋位点.半定量RT-PCR分析苦荞花期FtCHS空间表达模型表明,其表达量未成熟种子叶茎花根成熟种子,与苦荞芦丁含量的分布基本一致,具有组织特异性。     

苦荞查尔酮合成酶基因CHS的结构及花期不同组织表达量分析

采用同源克隆、染色体步移和RT-PCR技术,首次克隆到苦荞查尔酮合酶基因（CHS）的全长DNA序列和cDNA开放阅读框（ORF)序列. 序列分析表明,苦荞CHS DNA序列（GU172165）全长1 632 bp,含1个445 bp的内含子;cDNA编码区（HM852753）全长1 188 bp,编码395个氨基酸,命名为FtCHS. 生物信息学分析表明,FtCHS和推导的氨基酸序列与其它植物CHS基因同源率在95%以上,含有CHS多基因家族的标签序列（GFGPG）、活性位点、底物结合口袋位点和环化反应口袋位点. 半定量RT-PCR分析苦荞花期FtCHS空间表达模型表明,其表达量未成熟种子＞叶＞茎＞花＞根＞成熟种子,与苦荞芦丁含量的分布基本一致,具有组织特异性.     

血红密孔菌(Pycnoporussanguineus)漆酶基因的克隆与序列分析

为克隆血红密孔菌 (Pycnoporussanguineus)漆酶基因 ,根据真菌漆酶氨基酸序列保守区设计了 1对简并引物 .以血红密孔菌基因组DNA为模板 ,PCR扩增出长 12 2 7bp的漆酶基因片段 .以此序列为基础 ,通过 5′及 3′RACE技术克隆出漆酶全长cDNA序列 ,序列长为 190 2bp ,其 5′端和 3′端非编码区长分别为 5 1bp和 2 97bp ,开放阅读框长 15 5 4bp ,编码 5 18个氨基酸的蛋白 .该蛋白具有 4个铜离子结合区域 ,预测其相对分子量为 5 6 313 2 ,等电点为 5 5 9,其氨基酸序列与Pycnoporuscinnabarinus漆酶 (lcc3 2 )的同源性最高 ,为 96 % .以该cDNA编码区的两端序列为引物 ,PCR扩增得到漆酶的长度为 2 15 4bp的全长DNA序列 ,序列中包括 10个内含子序列 ,长为 5 2～ 70bp     

青花菜雄性不育相关基因BoDHAR的克隆与表达分析

以一个与甘蓝显性核不育相关的差异表达片段的序列为信息探针,通过在NCBI与TAIR网站数据库中进行同源EST序列搜索,经人工拼接、RT-PCR、PCR克隆与序列分析,获得了青花菜脱氢抗坏血酸还原酶DHARdehydroascorbatereductase基因的cDNA与DNA全长序列,命名为BoDHAR。并利用双链接头介导PCR的染色体步行技术(genomewalking)克隆了其上游644bp的5′端序列。所获的BoDHAR基因全长1486bp,存在两个内含子,DNA编码区序列633bp,编码210个氨基酸;序列分析表明BoDHAR与同源基因AT1G19570.1cDNA序列有82.3%的一致性,推导的氨基酸序列有79.6%的一致性;编码的水溶性蛋白存在多个磷酸化位点;5′端上游区存在明显的转录调控序列。半定量RT-PCR结果表明BoDHAR在可育系花蕾中的表达量明显高于不育系花蕾,在花药中的表达明显高于其它部位。     

Effects of MULTIFOLIATE-PINNA, AFILA, TENDRIL-LESS and UNIFOLIATA genes on leafblade architecture in Pisum sativum

In order to dissect the genetic regulation of leafblade morphogenesis, 16 genotypes of pea, constructed by combining the wild-type and mutant alleles of MFP, AF, TL and UNI genes, were quantitatively phenotyped. The morphological features of the three domains of leafblades of four genotypes, unknown earlier, were described. All the genotypes were found to differ in leafblade morphology. It was evident that MFP and TL functions acted as repressor of pinna ramification, in the distal domain. These functions, with and without interaction with UNI, also repressed the ramification of proximal pinnae in the absence of AF function. The expression of MFP and TL required UNI function. AF function was found to control leafblade architecture multifariously. The earlier identified role of AF as a repressor of UNI in the proximal domain was confirmed. Negative control of AF on the UNI-dependent pinna ramification in the distal domain was revealed. It was found that AF establishes a boundary between proximal and distal domains and activates formation of leaflet pinnae in the proximal domain.     

Presence of Rhizobium Etli bv . Phaseoli and Rhizobium Gallicum bv . Gallicum in Egyptian Soils

Seven bean rhizobial strains EBRI 2, 3, 21, 24, 26, 27 and 29 identified as Rhizobium etli, and EBRI 32 identified as Rhizobium gallicum, isolated from Egyptian soils and which nodulated Phaseolus vulgaris efficiently, were subjected to hybridization with a nifH probe in order to estimate the copy number of this gene. Seven strains (EBRI 2, 3, 21, 24, 26, 27 and 29) which were only able to nodulate Phaseolus vulgaris, contained three copies of the nifH gene, consistent with their identification as Rhizobium etli bv. phaseoli. Only one strain (EBRI 32) which nodulated both Phaseolus vulgaris and Leucaena leucocephala, had one copy of nifH gene. This confirmed the classification of this strain as Rhizobium gallicum bv. gallicum.     

Genome Analysis inNeurospora crassa;Cloning of Four Lociarginine-1(arg-1),methionine-6(met-6),unknown-7(un-7), andribosome production-1(rip-1) and Associated Chromosome Walking

We have cloned fourNeurospora crassagenes by complementation analysis. Cloned genes include thearginine-1(arg-1),methionine-6(met-6),unknown-7(un-7), andribosome production-1(rip-1) loci. Chromosome walks were initiated in ordered cosmid libraries from the cloned loci. A total of about 700 kb of theNeurosporagenome is covered in these walks.     

Characterisation of cell wall polysaccharides, arabinogalactans-proteins (AGPs) and phenolics of Cola nitida, Cola acuminata and Garcinia kola seeds

Many Cola plant species are endemic to West and Central Africa. Cola acuminata and Cola nitida are used as masticatory when fresh, while the dried nuts are used for beverages and pharmaceutical purposes in Europe and North America. Garcinia kola seeds, that serve as a substitute for the true kola nuts, are used in African traditional medicine for the treatment of various diseases, including colic, headache and liver cirrhosis. Seeds extracts of G. kola are also known for their anti-inflammatory, antimicrobial and antiviral properties. To gain information on the chemical properties of the kolas, we have isolated and analyzed cell wall polysaccharides, arabinogalactan-proteins and phenolic substances from the seeds of the three kola species. The sugar composition of cell wall material of C. acuminata, C. nitida and G. kola revealed that Gal (up to 30%), Ara, GalA and Glc as the predominant monosaccharides, representing approximately 90% by mol of the total hydrolysable sugar present in this material. In Ammonium oxalate cell wall fraction, GalA was found to be the major sugar present in all kola species. In the alkali-soluble fraction, there were significant differences in the level of Glc and Gal. The level of Glc was high in C. acuminata and C. nitida while the level of Gal and Xyl were high in C. nitida and G. cola. Isolation and quantification of arabinogalactan-proteins demonstrate that G. kola seeds contained four to eight times more of these proteoglycans than the seeds of the other two species. Finally, analysis of soluble phenolic substances shows that caffeine and catechin were largely represented in C. acumina and C. nitida seeds, with caffeine accounting for 50% of all soluble phenolics. These findings indicate that the three Kola seeds are highly enriched in pectins and proteoglycans and that C. acuminata and C. nitida can be used as a possible source of caffeine and catechin.     

中国黑粉菌属和利罗黑粉菌属

黑粉菌属是Roussel 1806年建立的,全世界记载有三百余种,主要寄生于禾本科,是经济作物及牧草的重要致病菌·长期以来,对黑粉菌的邢子使用过各种名称,如厚垣孢子,冬孢子及黑粉孢子等.本文采用黑粉孢子以区别锈菌的冬孢子. 芳’(1979)在《中国真菌总汇》中列出黑粉菌属五十种及一个变型.作者经过显微结构和超显微结构的研究,承认其中二十九种为正确名称,八种及一变型为异名,顶黑粉菌(Ustilago acrearus Berk.)由于错拼而被废弃.埃地黑粉菌(Ustilago emodensis Berk.)被转移至利罗粉菌属(Liroa).另有十一种黑粉菌因缺少标本留待今后订正.自1979年以后,杨信东(1983)增加黑粉菌属二种我国新纪录,K.范基和郭林(1986)描述一新种,四种新纪录.在本文中,作者描述一新种:鸢尾蒜黑粉(Ustilago ixiolirii Guo L) ,孢子堆生在蒴果内,不开裂,黑色,粉末状.黑粉孢子球形,近球形,稀椭圆形, 12.5-21×10-21μm,黑褐色,壁厚1-1.Sμm,纹饰脑状.是迄今生在石蒜科植物上唯一黑粉菌的种,其它几种黑粉菌均属条黑粉菌属.本文增加七种我国新纪录.共计四十九种,寄生于六科四十四属植物,主要是禾本科和蓼科.这仅是黑粉菌属研究的初步报告,在全国范围内大量采集黑粉菌标本后,作者相信会有更多新种和我国新纪录被发现.利罗黑粉菌属(Liroa)是从黑粉菌属(Ustaligo)分出的,此属为单种属.     

Nomenclatural changes resulting from the transfer of tropical African Sarcostemma to Cynanchum ( Apocynaceae : Asclepiadoideae )

Summary  Four species of tropical African Sarcostemma are transferred to Cynanchum together with two subspecies of S. viminale. In addition, Sarcostemma mulanjense is reduced to subspecific rank under C. viminale.     

The phylogenetic placement of Ostropales within Lecanoromycetes (Ascomycota) revisited

Results of molecular studies regarding the phylogenetic placement of the order Ostropales and related taxa within Lecanoromycetes were thus far inconclusive. Some analyses placed the order as sister to the rest of Lecanoromycetes, while others inferred a position nested within Lecanoromycetes. We assembled a data set of 101 species including sequences from nuLSU rDNA, mtSSU rDNA, and the nuclear protein-coding RPB1 for each species to examine the cause of incongruencies in previously published phylogenies. MP, minimum evolution, and Bayesian analyses were performed using the combined three-region data set and the single-gene data sets. The position of Ostropales nested in Lecanoromycetes is confirmed in all single-gene and concatenated analyses, and a placement as sister to the rest of Lecanoromycetes is significantly rejected using two independent methods of alternative topology testing. Acarosporales and related taxa (Acarosporaceae group) are basal in Lecanoromycetes. However, if the these basal taxa are excluded from the analyses, Ostropales appear to be sister to the rest of Lecanoromycetes, suggesting different ingroup rooting as the cause for deviating topologies in previously published phylogenies.     

转Cry1Ac/1Ab基因棉花对异色瓢虫生长发育及其捕食功能的影响

【目的】为探究转Cry1Ac/1Ab基因棉花对异色瓢虫生长发育及其捕食功能的影响。【方法】以转Cry1Ac/1Ab基因棉与其亲本常规棉为实验材料,利用取食不同棉花品种叶片的棉铃虫饲喂异色瓢虫幼虫。【结果】与常规亲本棉相比,取食饲喂转基因棉花叶片的初孵棉铃虫幼虫的异色瓢虫幼虫从1龄发育至化蛹期时间延长0.77 d,但差异不显著;除1龄幼虫体重增加(0.0773 mg)外,其余各龄期幼虫体重均有所下降,但差异均不显著;异色瓢虫1、2、3、4龄幼虫对初孵棉铃虫捕食量均随棉铃虫密度的增加而增加,捕食功能反应均符合HollingⅡ圆盘方程。【结论】转Cry1Ac/1Ab基因棉花对异色瓢虫生长发育无显著影响,饲喂取食转Cry1Ac/1Ab基因棉花的棉铃虫对异色瓢虫捕食功能无显著差异。     

Hydroxylation of steroids by Fusarium oxysporum, Exophiala jeanselmei and Ceratocystis paradoxa

The potential of Fusarium oxysporum var. cubense UAMH 9013 to perform steroid biotransformations was reinvestigated using single phase and pulse feed conditions. The following natural steroids served as substrates: dehydroepiandrosterone (1), pregnenolone (2), testosterone (3), progesterone (4), cortisone (5), prednisone (6), estrone (7) and sarsasapogenin (8). The results showed the possible presence of C-7 and C-15 hydroxylase enzymes. This hypothesis was explored using three synthetic androstanes: androstane-3,17-dione (9), androsta-4,6-diene-3,17-dione (10) and 3α,5α-cycloandrost-6-en-17-one (11). These fermentations of non-natural steroids showed that C-7 hydroxylation was as a result of that position being allylic. The evidence also pointed towards the presence of a C-15 hydroxylase enzyme.The eleven steroids were also fed to Exophialajeanselmei var. lecanii-corni UAMH 8783. The results showed that the fungus appears to have very active 5α and 14α-hydroxylase enzymes, and is also capable of carrying out allylic oxidations.Ceratocystis paradoxa UAMH 8784 was grown in the presence of the above-mentioned steroids. The results showed that monooxygenases which effect allylic hydroxylation and Baeyer–Villiger rearrangement were active. However, redox reactions predominated.