香蕉线条病毒病研究进展

香蕉是重要的经济作物和粮食作物,广泛种植于热带、亚热带地区.在我国的广东、福建、海南、广西和云南等地均有大面积种植,产销量一直居南方四大水果之冠.然而,近年来,许多病毒病害成为香蕉生产发展的主要限制因素,主要包括香蕉束顶病毒(banana bunchy top virus,BBTV)、香蕉线条病毒(banana streak virus,BSV)、香蕉苞片花叶病毒(banana bract mosaic virus,BBrMV)、黄瓜花叶病毒(cucumber mosaic virus,CMV)等引起的病害.     

特异扩增BBTV基因组Ⅲ、Ⅳ、Ⅰ编码区部分序列及其在检测中的应用

香蕉束顶病(BBTD)是香蕉植株的一种毁灭性病害,正在世界(包括中国)的许多香蕉种植区蔓延[1～7].其病原物为香蕉束顶病毒(Banana Bunchy Top Virus,BBTV),被列为我国第三类检疫对象.目前生产上主要采用培育脱毒组培苗来防治BBTD的发生,因此建立一种能快速、灵敏、特异地检测BBTV的方法就显得很重要.国内现在大多采用ELISA方法,但其灵敏度不够高,且需要制备特异性强的抗血清,否则较易出现假阳性.     

海南省香蕉枯萎病病原菌的分离鉴定及1号、4号小种的生物学特性

香蕉枯萎病是目前国内和世界很多地区香蕉种植区的一种毁灭性病害,目前尚未找到有效的防治方法.本文采集了海南省不同地区香蕉枯萎病的病原菌,通过经典病理学与分子生物学的方法进行了鉴定,此外还优化了快速检测的方法.结果表明,通过接种巴西香蕉和粉蕉进行致病性检测以及分离菌rDNA-ITS测序结果,明确鉴定出供试的香蕉分离菌株为4号生理小种,粉蕉分离菌株为1号生理小种.为了对1号、4号生理小种有更多的了解,本文还对1号、4号小种菌株生物学特性进行了测定,确定了影响菌丝生长的最佳条件.结果显示,pH 5时最适合1号、4号生理小种的菌丝生长,适合菌丝生长的温度为19～28℃之间,温度达到37℃时菌丝不能生长,碳源对菌丝的生长影响不大.这些方法和数据可以有利于田间香蕉枯萎病病害的检测和最终确定,生物学特性的测定可以为耕作技术的改进和农药的研发提供参考依据.     

组织培养香蕉试管苗的优势

随着组织培养技术的不断发展,组织培养法已成为生物科学研究的有力工具和农业经济生产的重要手段。尤其是在园艺植物上应用更为广泛泪前世界上大多数国家和地区都在开展植物组织培养的研究和应用,建立了诸如“基因工程公司”、“细胞技术公司”、“生命科学公司”、“试管苗公司”等专门机构,在农业上发挥着愈来愈重要的作用。香蕉试管苗,就是应用组织培养技术的原理和方法,采用香蕉的器官或组织,通过无菌操作,在适宜的人工培养基上进行培养,使其生长、增殖、分化形成的香蕉小植株。香蕉试管苗的推广应用,改变了过去一直采用的吸芽…     

香蕉束顶病毒研究进展

香蕉束顶病毒（Bananabunchytopvirus,BBTV）引起的香蕉束顶病（Ban。bu。bytoPdisease）是香蕉一种严重的病毒病害。迄今此病已普遍分布在世界许多产区,诸如：亚洲、非洲、澳大利亚、南太平洋一些岛屿以及美国的夏威夷等地区［‘－’l。在我国广东、广西、福建、云南等省的部分产区的发病率约占5—25％左右,严重地块已发展到毁灭性程度卜］。1987年Dale曾对世界香蕉种植的地理分布和BBTV的流行范围之间的关系、病株症状、病毒病原学、流行病学、病毒诊断方法以及病害的控制作过全面的综述【门。然而由于BBTV存在于寄主植物的韧皮…     

香蕉抗病基因工程

长期以来,香蕉生产遭受病害的严重威胁,制约了其发展,目前,随着转基因抗病香蕉基因工程技术的日趋成熟,为培育抗病香蕉品种开辟了新途径。     

芒果、香蕉采后病害生物防治的研究进展

芒果、香蕉采后主要病害为炭疽病、蒂腐病、冠腐病、黑腐病、黑星病.生物防治是当前芒果、香蕉采后病害控制的重要研究方向.概述了生物防治芒果、香蕉采后病害的方法,包括诱抗剂、植物提取物、拮抗微生物在芒果、香蕉采后病害防治上的研究与应用.     

基于多基因序列分析对尖孢镰孢菌古巴专化型（香蕉枯萎病菌）生理小种的鉴定

由尖孢镰孢菌古巴专化型Fusarium oxysporum f. sp. cubense, Foc引起的香蕉枯萎病是香蕉生产上的毁灭性病害,自1996年以来已对我国华南地区香蕉生产造成了严重危害。传统上香蕉枯萎病菌生理小种的鉴定主要采用人工接种鉴别寄主尔后测定病菌致病性的方法,但实验周期长,且受季节影响。以来自澳大利亚的香蕉枯萎病菌生理小种1号（BW1）、2号（Race 2）、3号（Race 3）以及亚热带4号（BW4）为对照,对分离自我国华南地区主要香蕉产区（广东、广西、海南、福建等省区）的14株香蕉枯萎病菌的单孢菌株进行致病性测定,并结合热带4号小种（TR4）和亚热带4号小种（ST4）的分子特异检测方法,确定其生理小种类型;同时,利用ITS、TEF-1α、IGS、histone H3、β-tubulin等 5个主要用于镰孢菌系统发育学研究的基因,研究不同地区不同来源的Foc菌株之间的亲缘关系及其与非病原尖孢镰孢菌的关系,并评价这5个基因在香蕉枯萎病菌生理小种鉴定上的应用价值。研究结果表明：（1）来源于我国华南地区的4号小种主要为热带4号小种;（2）TEF-1α、IGS、histone H3等3个基因片段能够将Foc中不同生理小种的菌株划分成不同的系统发育谱系,与致病性测定的结果具有对应关系,也能较好地反映尖孢镰孢菌种内菌株的亲缘关系,可用于香蕉枯萎病菌生理小种鉴定;（3）我国Foc 1号生理小种的遗传多样性高于4号生理小种,Foc 1号生理小种的菌系与来自香蕉果实上的非病原尖孢镰孢菌的亲缘关系比其与Foc 4号生理小种的菌系的亲缘关系更近。     

香蕉束顶病毒研究进展

香蕉束顶病毒(Bananabunchytopvirus,BBTV)是引起香蕉束顶病害(Bananabunchytopdisease,BBTD)的病毒,它严重地危害了香蕉的生产。综述了近年来香蕉束顶病毒的分离提纯方法,株系划分以及分类地位,较为全面的介绍了BBTV病毒基因组分结构和各组分编码蛋白的功能等,并提出了目前需要进一步澄清的问题。     

新型杀菌剂啶氧菌酯对香蕉叶斑病的防治效果

【目的】香蕉叶斑病是香蕉产业的重要病害,化学防治仍然是当前最为有效的防治手段。于2015—2016年连续2年开展香蕉叶斑病的田间化学防治试验,为生产上推广应用新型杀菌剂啶氧菌酯提供依据。【方法】试验设22.5%啶氧菌酯悬浮剂125、150和187.5 mg·kg~(-1),对照药剂250 g·L-1吡唑醚菌酯乳油125 mg·kg~(-1),以及空白对照共5个处理,3次药后第12或13天调查正常叶数、病叶数及病级,计算平均病指及平均防效。【结果】22.5%啶氧菌酯悬浮剂(有效成分用量125、150和187.5 mg·kg~(-1))2015年的防治效果分别为64.70%、68.16%和71.29%,2016年防治效果分别为68.44%、72.36%和76.29%。此外,在试验期间香蕉嫩叶未见药害现象,叶片生长均正常。【结论】22.5%啶氧菌酯悬浮剂是防治香蕉叶斑病的优良药剂,对香蕉比较安全,值得在香蕉产区推广应用。     

香蕉束顶病毒基因克隆和序列分析

对香蕉束顶病毒（ＢＢＴＶ）中国分离株ＤＮＡ组份Ｉ（ＤＮＡ－１）、外壳蛋白（ＣＰ）和运转蛋白（ＭＰ）基因进行了克隆和序列分析。ＢＢＴＶＤＮＡ－１含有１１０３个核苷酸,与南太平洋和亚洲分离株分别有８７％－８８％ ９６．９－９８％的核苷酸序列同源性。由ＤＮＡ－１编码的复制酶含有１８６个在酸残基。与南太平洋和亚洲分离株分别有８４．４％－９５．８％和９７．６％、９８．０％的氨基酸序列同源性。外壳蛋白基因由５     

Two Circular Single-stranded DNAs Associated with Banana Bunchy Top Virus

Nucleic acids extracted from partially purified banana bunchy top virus (BBTV) consisted of 20 Kb DNA, 0.9-1.1 Kb DNA and 0.3 Kb RNA. Partially purified BBTV preparations predigested with DNase and RNase before particle disruption and nucleic acid isolation yielded only the 0.9-1.1 Kb DNA, but no corresponding nucleic acid band was obtained in total nucleic acid isolated from healthy banana tissue. Analysis of two BBTV cDNA clones showed that clone 1 consisted of 287 nucleotides and clone 2 contained a 1.0 Kb DNA insert. Clone 1 is not part of clone 2. When two pairs of primers, each pair in opposite orientation were used to amplify BBTV DNA by PCR using the total DNA from diseased banana tissues or DNA encapsidated in BBTV particle as the template, a DNA product of 1.1 Kb was generated by both, results indicating that the BBTV DNAs are circular. Additional results suggested that BBTV contained at least two circular ssDNAs designated BBTV essDNA I (containing clone 1 nucleotide sequence) and BBTV, cssDNA II (containing clone 2 nucleotide sequence).     

Two Circular Single-stranded DNAs Associated with Banana Bunchy Top Virus

Nucleic acids extracted from partially purified banana bunchy top virus (BBTV) consisted of 20 Kb DNA, 0.9–1.1 Kb DNA and 0.3 Kb RNA. Partially purified BBTV preparations predigested with DNase and RNase before particle disruption and nucleic acid isolation yielded only the 0.9–1.1 Kb DNA, but no corresponding nucleic acid band was obtained in total nucleic acid isolated from healthy banana tissue. Analysis of two BBTV cDNA clones showed that clone 1 consisted of 287 nucleotides and clone 2 contained a 1.0 Kb DNA insert. Clone 1 is not part of clone 2. When two pairs of primers, each pair in opposite orientation were used to amplify BBTV DNA by PCR using the total DNA from diseased banana tissues or DNA encapsidated in BBTV particle as the template, a DNA product of 1.1 Kb was generated by both, results indicating that the BBTV DNAs are circular. Additional results suggested that BBTV contained at least two circular ssDNAs designated BBTV cssDNA I (containing clone 1 nucleotide sequence) and BBTV cssDNA II (containing clone 2 nucleotide sequence).     

香蕉束顶病毒DNA组分2、3的启动子区的组织特异性分析

香蕉束顶病毒（BBTV）基因组至少由6个大小约为1.0-1.1kb的单链环状DNA组分所组成,每一个DNA组分包含编码区与非编码区。本文在前人的研究基础上进一步了解BBTV DNA组分启动子的功能。首先根据BBTV 海南分离物的全序列,通过常规PCR扩增出长为540bp的 BBTV DNA3组分启动子序列BV3.1,同时通过重叠PCR扩增出646bp的DNA2与DNA3组分非编码区拼接的重组启动子序列BV23,分别替代pBI121 35S启动子序列与gus基因进行融合,构建植物表达载体pBIBV3.1、pBIBV23。农杆菌介导转化获得的pBIBV3.1转基因烟草经GUS化学组织染色后,在其叶片的叶脉处检测到微弱的GUS活性,证实了DNA3组分的韧皮部特异表达活性;而pBIBV23转基因烟草,其叶片经GUS组织化学染色后,在叶肉、叶缘及一些叶脉上检测到弱GUS活性,这表明由BV23驱动的gus基因在烟草中类似于组成型表达,则DNA2组分转录方式可能有异于DNA3组分。     

Molecular Characterization of a Strain of Squash Leaf Curl China Virus from the Philippines

The complete nucleotide sequence of infectious cloned DNA components (A and B) of the causal agent of squash leaf curl disease in the Philippines was determined. DNA‐A and DNA‐B comprise 2739 and 2705 nucleotides, respectively; the common region is 174 bases in length. Five ORFs were found in DNA‐A and two in DNA‐B. Partial dimeric clones containing DNA‐A and DNA‐B, constructed in a binary vector and transformed into Agrobacterium tumefaciens, induced systemic infection in agro‐inoculated pumpkin plants (Cucurbita moschata). The total DNA‐A sequence was most closely related to that of Squash leaf curl China virus (SLCCNV) (88% identity), although the existence of B component of SLCCNV has not been reported. The deduced coat protein was like that of SLCCNV (98% amino acid sequence identity) and the Philippines virus has low sequence identity to Squash leaf curl virus (SLCV) and Squash mild leaf curl virus (SMLCV) (63 and 64% total nucleotide sequence identities, respectively). From these results, we propose that the Philippines virus be designated Squash leaf curl China virus‐[Philippines] (SLCCNV‐[PH]).     

Molecular comparative analysis of component 1 (DNA-R) of an Egyptian isolate of banana bunchy top nanovirus isolated from banana aphid (Pentalonia nigronervosa)

We are reporting a molecular comparative analysis of component 1 BBTV-DNA-R of an Egyptian isolate of (BBTV) and 30 different geographical isolates. DNA was extracted from BBTV-infected adult banana aphids collected from El-Qalubia Governorate, Egypt. Using specific primers the BBTV-DNA-R was amplified, cloned into a prokaryote vector, sequenced and a molecular comparative analysis of BBTV-DNA-R of this study and some overseas isolates of BBTV-infected banana plants was determined. Results showed that the component 1 consists of 1108 nts and contains a sequence of 69 nts representing the CR-SL of 31 nts. A CR-M (90 nts) at the position (972–1062) characterized with GC-rich sequence from nts 76 to 90 (average of 80% G + C) was found. Alignment results of BBTV DNA-R confirmed the presence of a number of conserved regions in all isolates. Large ORF of 861 nts at position 102 to 962 in the virion sense were detected. The predicted protein of this ORF consisted of 286 amino acids and had a molecular weight of 33.8 kDa. The DNA-phylogenetic analysis showed a percent identity of 98.0 and 97.9 between BBTV DNA-R and isolates of Pakistan (isolate TJ1) and Australia (isolate V1), respectively. The similarities between the gene product of Egyptian BBTV DNA-R and the 30 overseas isolates ranged from 93.7 to 99.0%. Differences in phylogenetic trees based on the entire sequence of BBTV DNA-R, CR-M and amino acid sequences confirmed the existence of two taxonomic groups of BBTV and the Egyptian isolate belongs to the south pacific group.     

香蕉束顶病毒复制酶基因克隆及转基因表达

以广州市郊获得的香蕉束项病毒(BBTV)的DNA为模板,进行PCR扩增得到香蕉束项病毒复制酶基因的1.1 kb DNA.所获得的DNA序列与澳大利亚的BBTV序列的同源性达90%,这部分序列编码香蕉束顶病毒复制酶基因的羧基端.将改造的BBTV复制酶基因克隆到pBll21的CaMV 35S和NOS终止序列之间,构建表达载体,并采用基因枪轰击香蕉试管苗生长点组织的方法,经PCR检测和Westem blot分析,获得4株具有BBTV复制酶基因整合表达的To代转基因香蕉.转基因植株的抗病性正在检测之中.     

Association of a Distinct Begomovirus and a Betasatellite with Leaf Curl Symptoms in Pedilanthus tithymaloides

Pedilanthus tithymaloides (Redbird flower) is an ornamental shrub that occasionally exhibits leaf curl and enation symptoms in Pakistan. Symptoms were shown to be associated with a monopartite begomovirus and a betasatellite. The complete nucleotide sequence of the begomovirus was found to be 2764 nucleotides in length and have the highest nucleotide sequence identity to a begomovirus previously isolated from tomato (90.3% nucleotide sequence identity), followed by Radish leaf curl virus (86.3%). The complete betasatellite sequence was determined to be 1358 nucleotides in length and has the highest sequence identity (97%) with Tobacco leaf curl betasatellite . The analysis shows the begomovirus associated with leaf curl disease of Pedilanthus to be a distinct and previously unreported begomovirus for which the name Pedilanthus leaf curl virus (PedLCV) is proposed. This virus is one of an increasing number of monopartite begomoviruses shown to be associated with a betasatellite.     

Arrangement of a highly repeated DNA sequence in the genome and chromatin of the African green monkey.

The DNA of the African green monkey contains three components that are distinguishable by the kinetics of reassociation. The rapidly reassociating component represents about 20% of the total DNA and is composed almost entirely of a sequence (AGMr(HindIII)-1) which is repeated 6.8 x 10(6) times. The majority of the AGMr(HindIII)-1 sequences are organized in long tandem repeats of a segment of 172 base pairs in length. However, a fraction of the AGMr (HindIII)-1 sequences is interspersed with another 37% of the genome. The structure of the chromatin containing the AGMr-(HindIII)-1 sequence is indistinguishable from that containing total DNA. Furthermore, there is nothing inherent in the nucleotide sequence of AGMr(HindIII)-1 which specifies a unique location for nucleosomes.     

Limited permutations of the nucleotide sequence in bacteriophage T1 DNA

In the mature DNA molecules of bacteriophage T1 three different nucleotide sequence arrangements are found, differing from each other by circular permutations through 6% or 12% of the total length. Using a short region of non-homology between T1⁺ and T1Ds DNAs as a sequence marker, we have measured the positions of this marker with respect to the ends of the individual component strands in T1⁺: T1Ds heteroduplexes. The frequency distribution shows three peaks, at 37%, 43% and 49% of the total length, measured from the free end in molecules, one of whose ends is identified by being bound to the phage ghost. Since the distance between peaks is equal to the length of the terminally repeated segment of nucleotide sequence in T1DNA (6.1% of the molecular length), this finding suggests that T1 DNA is matured by the headful mode, commencing from a particular initiation site within the nucleotide sequence and proceeding with the maturation of only three consecutive headfuls (or sometimes two, and rarely four). The relative frequency of occurrence at the three principal positions, approximately 0.4:0.4:0.2, respectively, suggests that the maturation initiation site lies at the ghost-bound end in the permutation identified with the 37% position of the Ds non-homology.