甘蔗黄叶病及其病原分子生物学研究进展

甘蔗黄叶病是由甘蔗黄叶病毒(Sugarcane yellow leaf virus,SCYLV)引起的一种病毒病害,在全球主要甘蔗种植国家或地区普遍发生,危害日益严重。SCYLV经甘蔗蚜虫以持久性方式传播,在寄主植株内主要侵染韧皮部组织。该病毒起源于黄症病毒科属间基因重组,被国际病毒分类命名委员会确认为黄症病毒科马铃薯卷叶病毒属成员。文章综述了甘蔗黄叶病毒生物学特征、病害发生和危害、病原鉴定和检测、分子进化和遗传多样性、基因组结构和基因功能以及抗病转基因等方面研究进展,并对甘蔗黄叶病抗病育种和防治措施作了讨论。     

Symptom expression of yellow leaf disease in sugarcane cultivars with different degrees of infection by Sugarcane yellow leaf virus

Sugarcane yellow leaf virus (ScYLV) is present in many sugarcane growing areas of the world. It is suspected to cause yellow leaf disease (formerly called YLS, yellow leaf syndrome) of sugarcane. This study investigated symptom expression in a selection of cultivars classified into three groups; ScYLV-susceptible/infected, ScYLV-resistant and intermediately infected cultivars grown in plantation fields in the islands of Hawaii. Incidence of yellow leaf symptoms was correlated, though not tightly, to the presence of ScYLV. The correlation is based on two factors: (i) only ScYLV-infected cultivars (from both susceptible and intermediate groups) showed severe symptom expression, and (ii) ScYLV-infected plants had four times higher symptom incidence than virus-free plants of the same cultivar. The yellow leaf symptom expression fluctuated, peaking at 200, 350, 500 and 600 days after planting. These symptom peaks were correlated with an increase of ScYLV content in the intermediately infected group of cultivars. No nutritional, environmental or field factor could be identified which clearly influenced symptom expression. It is speculated that the symptom expression is elicited by assimilate backup in the stalks and that the fluctuation of symptom expression is caused by the growth rhythm of mature sugarcane stalks.     

Transmission of a sugarcane yellow leaf phytoplasma by the delphacid planthopper Saccharosydne saccharivora, a new vector of sugarcane yellow leaf syndrome

During surveys of sugarcane fields in western and central Cuba from December 2001 to March 2003, the delphacid planthopper Saccharosydne saccharivora was the most prevalent of the Auchenorrhyncha fauna surveyed. Individuals of S. saccharivora collected tested positive for the sugarcane yellow leaf phytoplasma (SCYLP). Saccharosydne saccharivora were reared in cages and used for experimental transmission studies of SCYLP. The S. saccharivora were given acquisition-access feeds of 72 h on SCYLP-infected canes collected from the field followed by an inoculation-access period of 15 days on healthy sugarcane seedlings. Symptoms of yellow leaf syndrome developed on 24 out of 36 plants, 7–12 months postinoculation. None of the 36 healthy seedlings that were inoculated with S. saccharivora fed on phytoplasma-free sugarcane developed symptoms. All phytoplasma-positive sugarcane and S. saccharivora samples showed identical RFLP patterns and had 99·89% similarity in their 16S/23S spacer-region sequences, but only 92·6–93·6% similarity with other phytoplasmas. Sequences were deposited with GenBank [accession numbers: AY725237 ( S. saccharivora ) and AY257548 (sugarcane)]. Phylogenetic analysis suggested that the phytoplasmas from sugarcane and S. saccharivora are putative members of a new 16Sr phytoplasma group. This is the first report of vector transmission of a phytoplasma associated with sugarcane yellow leaf syndrome and the first time that S. saccharivora has been shown to vector a phytoplasma.     

番茄黄化曲叶病毒与番茄褪绿病毒复合侵染对番茄黄化曲叶病毒传播的影响

番茄黄化曲叶病毒(tomato yellow leaf curl virus, TYLCV)是一种由烟粉虱传播的单链环状DNA病毒, 在田间可与多种病毒发生复合侵染, 如番茄褪绿病毒(tomato chlorosis virus, ToCV)等?本文对比了TYLCV单独侵染和TYLCV与ToCV复合侵染对烟粉虱获取和传播TYLCV的影响?结果表明, 与取食TYLCV单独侵染的番茄相比, 取食复合侵染番茄的烟粉虱对TYLCV的传毒率显著提高, 且番茄植株和烟粉虱体内TYLCV的病毒积累量也显著提高?试验结果说明复合侵染会提高烟粉虱的传毒率, 促进TYLCV的发生与流行?     

Detection and typing of tomato yellow leaf curl viruses

The denomination Tomato yellow leaf curl virus (TYLCV) comprises several viruses that cause severe damage to tomato crops in warm and temperate regions worldwide. TYLCV viruses are widespread in the Mediterranean Basin, in which two species have been reported: Tomato yellow leaf curl Sardinia virus (TYLCSV) and Tomato yellow leaf curl virus (TYLCV, previously TYLCV-Is). The availability of methods convenient for the diagnosis of these viruses is essential. We have investigated several alternatives for reliable detection and differentiation of TYLCSV and TYLCV. Triple-antibody sandwich enzyme-linked immunosorbent assay (TAS-ELISA) proved to be very useful for large-scale diagnosis in field situations, but lacked discriminating capacity and sensitivity in the stages of infection in which low virus titre is present. The DNA-based methods are suited to laboratory operations and plant disease clinics, where accuracy of detection and discrimination of viruses is required. Polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) was the most reliable method to discriminate between TYLCSV and TYLCV, but is not suited to high sample turnover. For large-scale testing, tissue print hybridization assay provides a reliable and sensitive alternative to PCR.     

Cultural management of tomato yellow leaf curl disease in Cyprus

Cultural management of tomato yellow leaf curl virus (TYLCV) by roguing old, overwintered tomato crops was investigated in a small, isolated production centre in the southern coastal zone of Cyprus. Control measures were applied on an area-wide basis during March-April, before emergence of adult whitefly ( Bemisia tabaci Genn.) vectors, for three consecutive years (1980-82). In all three years, primary virus spread to spring plantings was almost completely prevented, while further secondary spread to summer plantings was kept below 5%, compared with 40-50% in 1978 and 1979. When control measures were discontinued disease incidence increased to 15% in 1983 and 40% in 1984. The disease could also be controlled by early transplanting in the spring or by late transplanting in the autumn. Several cultural practices were evaluated to prevent or reduce infection of seedlings before transplanting. Seedlings produced in the summer in a coastal region were infected by TYLCV (virus incidence 7-42%), with a higher incidence in seedbeds situated near infected tomato crops. Infection was significantly reduced by covering seedbeds with an insect-proof tunnel, and was completely prevented by producing seedlings in inland areas, where no commercial tomato crops were grown. The use of healthy transplants delayed subsequent disease development in the field but had little effect on final disease incidence.     

Current situation of Tomato yellow leaf curl virus in Portugal

The name Tomato yellow leaf curl virus (TYLCV) has been applied to a group of virus species of the genus Begomovirus in the family Geminiviridae that cause a similar tomato disease worldwide. In 1995, TYLCV was first reported in Algarve (southern Portugal) as responsible for an epidemic outbreak of a severe tomato disease. Molecular data have shown that this Portuguese TYLCV isolate was distinct from those previously reported in Europe, as it belonged to the TYLCV-Israel species ¹ . Since then, TYLCV epidemics have occurred annually, being a limiting factor mainly for autumn/winter glasshouse tomato crops. In 1998, TYLCV was also found associated with the emergence of a novel disease of Phaseolus vulgaris in Algarve. The affected bean plants were severely stunted and gave no marketable yield. However, the disease occurs only sporadically, even in conditions of high TYLCV infection pressure. Recently, Tomato chlorosis virus (ToCV), a whitefly-transmitted bipartite closterovirus (genus Crinivirus , family Closteroviridae ), was found associated with an unusual tomato yellow leaf syndrome, in single or mixed infection with TYLCV. The impact of this new pathosystem on tomato production has yet to be determined. Surveys are in progress in mixed cropping systems infested with whiteflies. So far, TYLCV and ToCV diseases are limited to the Algarve region.     

河北省番茄黄化曲叶病毒病的分子鉴定初报

双生病毒是一类具有孪生颗粒形态的植物单链病毒,目前双生病毒病害已在多个国家和地区的作物上造成严重危害。近年来,我国多省报道作物上有这类病毒的发生,且有逐年加重和扩散的趋势。根据危害番茄的双生病毒DNA A序列保守区设计简并引物,对2009年4月采自河北省魏县表现叶片黄化、曲叶症状的4个番茄样品进行PCR检测,均为双生病毒阳性,对样品的扩增片断进行了克隆测序,经序列比对分析,与番茄黄化曲叶病毒（Tomato leaf curl virus,TYLCV）山东分离物（FJ646611.1）序列相似性为99.25%~99.55%,说明河北番茄黄化曲叶病由TYLCV引起。     

Detection of Tomato yellow leaf curl Sardinia virus in Tunisia

Surveys were undertaken for tomato yellow leaf curl viruses in the main Tunisian tomato-growing areas in fields and plastic houses. Symptoms included yellowing, leaf curling and stunting. Collected samples were submitted to molecular analysis using two approaches: (1) hybridization tests with two DNA probes corresponding to an intergenic region derived from a cloned dimer of an Egyptian full-length TYLCV and to the coat region of a Tunisian TYLCV isolate; (2) PCR amplification coupled to RFLP allowing both identification and clustering of Tunisian isolates. Only Tomato yellow leaf curl Sardinia virus was detected in Tunisia.     

Breeding tomatoes for resistance to tomato yellow leaf curl begomovirus

Tomato yellow leaf curl begomovirus (TYLCV) can be devastating to tomato crops in tropical and subtropical regions. The development of resistant cultivars is the best option for the control of TYLCV. The TYLCV-resistance level of a new breeding line, TY172, alongside that of commercial cultivars known to be resistant to the virus, was evaluated in a field test by comparing the yield performance of inoculated plants with that of uninoculated plants of the same line or cultivar. There were substantial differences among the different entries tested in the extent of yield loss relative to the corresponding uninoculated control plants. This comparison between inoculated and uninoculated plants of the same entry provides a quantitative assay for resistance level. All resistant commercial cultivars tested developed different levels of disease symptoms. Only line TY172 showed no symptoms of the disease. A low level of viral DNA was detected in infected TY172, showing that it is a symptomless carrier of TYLCV. When TY172 was crossed with susceptible lines, the hybrids exhibited milder symptoms than the susceptible parent, yet higher than that of TY172, suggesting a partial dominance for TY172 resistance. Upon inoculation of F₂ populations, the amount of symptomless individuals appeared in a ratio approximating 7:64. This suggests that at least three genes appear to account for the resistance.     

柑桔黄斑落叶病病原菌鉴定

正柑桔(Citrus reticulata Blanco)是著名的热带、亚热带水果。位于我国南疆的广西,属于亚热带季风气候,是种植柑桔最适宜区或适宜区,柑桔也是广西一大优势水果产业,是广西出口东盟的主要农产品。柑桔黄斑落叶病是近年来在我国广西地区发生的一种新病害。该病害主要危害秋梢和春梢叶片,形成不规则黄斑,严重的可造成整个叶片脱     

Tomato yellow leaf curl virus: host range and virus-vector relationships

The Jordanian isolate of tomato yellow leaf curl virus has a narrow host range restricted to a few solanaceous plants. Severe symptoms developed on tomatoes and Datura stramonium , whereas Nicotiana glutinosa and N. tabacum cvs Samsun and Havana 423 were infected without showing symptoms. The whitefly Bemisa tabaci is an efficient vector; a single whitefly was able to transmit the virus. The minimum acquisition and inoculation feeding periods were 60 and 30 min, respectively, and the latent period was 20–24 h. The virus was retained by Bemisia tabaci for 11 days. The host range and virus vector relationships of the Jordanian isolate are reported for the first time.     

韭菜黄叶病病原细菌的鉴定

在桂中地区韭菜上发现一种由细菌引起的病害,该病主要发生在外叶,表现出纵向半片叶或全片叶变黄。从叶片的病组织中分离到具有致病性的杆状细菌,将分离的病原菌接种于健康的韭菜上,表现与田间一致的症状。接种发病的病斑与田间标本上分离的病原菌菌落形态完全相同。根据菌株的培养性状、常规生理生化特性及透射电镜下菌体形态观察结果,将病原菌鉴定为草生欧文氏菌菠萝变种(Erwinia herbicolavar.ananas)。     

侵染广西番茄的番茄黄化曲叶病毒的分子鉴定

番茄黄化曲叶病毒病是番茄生产上的毁灭性病害,严重影响番茄产量及品质。2019年从广西百色市采集疑似番茄黄化曲叶病叶片样品,采用滚环扩增(RCA)及基因克隆等方法,获得了4个烟粉虱传双生病毒的全基因序列,4个分离物的全长序列分别为2 776、2 781、2 752、2 781 bp,均编码6个开放阅读框;核苷酸相似性比较发现,4个分离物彼此间的相似性均在90%以上,与已报道的番茄黄化曲叶病毒Tomato yellow leaf curl virus (TYLCV)各分离物间的相似性也在91%以上;系统进化分析表明,TYLCV广西分离物BS-2和BS-4与TYLCV-Hunan、BS-1和BS-3与TYLCV-YN6553等分离物处于独立的小分支,说明TYLCV广西分离物与TYLCV-Hunan、TYLCV-YN6553具有较近的亲缘关系。本研究首次报道TYLCV在广西发生。     

2009年河北省番茄黄化曲叶病毒病发生危害和分布

2009年,河北省主要番茄种植区发生一种疑似番茄黄化曲叶病毒病的新病害。本文在田间调查、取样的基础上,采用酶联免疫吸附测定(ID-ELISA)方法对病原进行鉴定,明确了番茄黄化曲叶病在河北省的分布与危害:至2009年9月,番茄黄化曲叶病毒病已在河北省南部的邯郸、邢台和石家庄,中部的保定和衡水等地严重发生,河北省中北部廊坊、沧州南部靠近衡水的部分地市亦零星发生此病害,应用TYLCV检测试剂盒检测的29个品种58个标样A405值达到2.1～7.5。唐山、承德地区尚未发现番茄黄化曲叶病的危害,4个县市9个番茄品种检测结果均呈阴性。     

北京地区番茄黄化曲叶病毒病的鉴定及防治对策

番茄黄化曲叶病毒病是一种由烟粉虱传播的病毒病,给番茄生产造成严重威胁。2009年在北京郊区调查时发现部分保护地种植的番茄植株表现典型黄化曲叶症状。通过提取典型症状样品总DNA利用粉虱传双生病毒检测简并引物PA/PB,进行PCR扩增到541bp的特异条带。通过测序和核苷酸序列比对表明该序列与番茄黄化曲叶病毒序列相似性最高为99%。分子检测结果表明北京郊区部分保护地种植的番茄已被烟粉虱传播的番茄黄化曲叶病毒侵染危害。     

Tomato yellow leaf curl virus effects on chloroplast biogenesis and cellular structure

Tomato yellow leaf curl virus (TYLCV) is a begomovirus causing significant crop losses in tomatoes worldwide. TYLCV infection has several defined symptoms that are characterized at the whole plant level that include yellowing and curling of the leaves, stunting and growth retardation, and severe yield losses that can reach 100%. While the etiology and phenotypic symptoms of TYLCV infection were characterized, little is known about the cellular basis of these symptoms. Our goal in the present study is to describe some of the microscopic features of the disease while comparing TYLCV-susceptible and resistant near-isogenic tomato plants under infection. We depict the cellular changes that occur in susceptible line as a result of TYLCV infection that include: epidermal cell sizes reduction, accompanied with modifications in cell shape; reduction in plastid number, plastid size and chlorophyll content; diminished expression genes encoding LHC, Rub L, CAB4, and CAB8 and a surge in plasmodesmata number after infection. We conclude that TYLCV affect many targets or that it has a very critical target in cellular development.