Use of netting and whitewash spray to protect papaya plants against Nivun Haamir (NH)-dieback disease

A disease causing the decline of papaya (Carica papaya) plants was noticed in the Jordan Valley in 1982. The disease caused severe yellowing of the upper leaves and dieback of the apex. The disease was not transmissible mechanically and continued efforts to associate viruses and viroids have so far been unsuccessful. Epidemiological observations suggested that the disease is airborne and probably caused by a mollicute. In order to test this hypothesis, plots were covered by an insect-proof net and with white nets providing 15, 30 and 50% shade. The 30%, 50% and insect-proof nettings provided complete protection, and the 15% netting reduced disease incidence to <2%, compared with>37% in the uncovered control plot. Spraying plants at weekly intervals with a commercial whitewash solution was also found to be effective in reducing disease incidence. The advantages of using a range of netting field treatments as a simple means for obtaining information on the epidemiological nature of a new disease are described.     

番木瓜遗传改良研究进展

就番木瓜的株性和抗病性等性状遗传、杂交育种、人工诱变育种、离体培养、分子标记及基因工程育种等现代生物新技术育种方面的研究进展及相关问题进行了评述,重点综述了番木瓜株性遗传、远缘杂交与胚胎学、基因工程在抗环斑型花叶病毒病(PRSV)品种培育及分子标记辅助选择育种应用的研究进展,并分析了今后番木瓜育种的发展趋势。     

番木瓜环斑病毒株系的分子生物学方法鉴定

 以PRSV株系特异性引物对PRSV的PRSV126(PRSV日本分离物)、Ys、Vb和Sm等株系进行RT-PCR方法鉴定,引物PR21/PR22能把Ys从Vb和Sm中鉴定出来,PR300/PR301则能把Vb从Ys、Sm和PRSV126中鉴定出来;用限制性内切酶Hae Ⅱ、Sau3A I和Hinf I对PRSV的PRSV126、Ys、Vb和Sm等株系进行单酶切RT-PCR-RFLP分析,Hinf I能把PRSV126与Ys、Vb和Sm鉴别开来,Sau3A I能把Ys与Vb和Sm鉴别开来,Hae Ⅱ则能把Ys与PRSV126、Vb和Sm鉴别开来;以P1/P2为引物,对Vb、Ys和Sm株系进行RT-PCR-RFLP-SSCP分析,结果能一次把三者较好地区别开来。     

Purification and some properties of Papaya meleira virus, a novel virus infecting papayas in Brazil

'Meleira', or 'sticky disease', is currently the most damaging papaya disease in the mid-eastern Brazilian growing regions. Consistent disease transmission via latex injection, presence of similar isometric particles in the laticiferous vessels of diseased plants, and detection of double-stranded DNA in naturally and experimentally infected papaya trees suggest that a virus is the causal agent. Conclusive evidence for viral aetiology was previously lacking, mostly because every attempt to purify the putative virus from infected papayas had failed. Following the successful purification and partial characterization of the meleira virus, healthy papaya seedlings injected with purified virus particles later developed typical symptoms of the disease. Negatively stained, isometric, full and 'empty' purified virus particles measured 42 and 38 nm, respectively. The viral genome was a single dsRNA molecule of about 12 kbp. Several capsid proteins, ranging in size from 14·4 to 45 kDa, were consistently revealed by PAGE. Papaya meleira virus (PMeV) appears to represent a novel group of viruses, with no known similar counterpart among known plant-, vertebrate-, invertebrate- or prokaryote-infecting viruses.     

番木瓜环斑病毒弱毒株的构建及分析

弱毒株是利用交叉保护防治植物病毒病害的关键限制因子。通过对马铃薯Y病毒属病毒(强、弱毒株)的全长及部分氨基酸序列比对分析,筛选出在亲本强毒株PRSV-LM的P1和HC-Pro基因上的8个可能与致病性相关的氨基酸突变位点(I_(309)、K_(481)、K_(598)、R_(728)、F_(753)、L_(754)、N_(787)和D_(944))。采用定点突变和Gibson拼接法,分别成功构建了4种含有2个突变位点(L_(754)和N_(787))、4个突变位点(I_(309)、K_(481)、F_(753)和D_(944))、6个突变位点(I_(309)、K_(481)、F_(753)、L_(754)、N_(787)和D_(944))和8个突变位点(I_(309)、K_(481)、K_(598)、R_(728)、F_(753)、L_(754)、N_(787)和D_(944))的PRSV-LM全长c DNA侵染性克隆突变体:p Gprsvm~2、p Gprsvm4、p Gprsvm6和p Gprsvm8。经人工接种、病症观察和RT-PCR检测,4种突变体克隆均能系统性侵染非转基因番木瓜植株,除克隆p Gprsvm~2的病症表现与亲本强毒株PRSV-LM一致外,其他多位点突变的克隆p Gprsvm4、p Gprsvm6和p Gprsvm8在番木瓜植株上的病症严重程度出现依次减弱,而克隆p Gprsvm8的病症最弱,且延迟2周发病。本实验初步验证了6个氨基酸突变位点(I_(309)→S、K_(481)→Q、K_(598)→E、F_(753)→L、R_(728)→I和D_(944)→N)与PRSV-LM的病症表现及致病力相关,其中K_(598)→E和R_(728)→I可能是影响致病性的关键位点。     

海南番木瓜PRSV和PLDMV病毒发生情况及分子鉴定

对海南昌江、乐东、澄迈、文昌和三亚5个主要番木瓜产区的病毒病发生情况进行调查。对76份番木瓜疑似病毒样品进行检测,检测结果表明海南番木瓜病毒病主要有2种：由番木瓜环斑病毒（Papaya ring spot virus,PRSV）引起的番木瓜环斑病毒病和由番木瓜畸形花叶病毒（Papaya leaf-distortion mosaic virus,PLDMV）引起的番木瓜畸形花叶病毒病。其中,番木瓜环斑病毒病最为常见,检出率达77.63％,是影响海南地区番木瓜产业健康发展的主要病毒病;畸形花叶病毒病发生率很低,仅检出2例番木瓜畸形花叶病毒病样品。以PRSV病毒基因组P1和Hc-Pro作为靶标基因进行分子克隆、测序,结果表明海南地区PRSV病毒可明显分为2个株系：HN-PRSV-1株系和HN-PRSV-2株系。此外,基于CP基因的系统进化树分析结果表明在海南新发现的PLDMV病毒可能源于台湾和日本。本研究可为开展番木瓜花叶病的防控和创制抗花叶病毒番木瓜新种质提供数据参考。     

Papaya yellow leaf curl virus: A newly identified begomovirus infecting Carica papaya L. from the Indian Subcontinent

India is one of the world’s largest producers of papaya. Viruses, mainly begomoviruses and potyviruses, cause a significant loss in papaya production. The study described here has identified a new species of begomovirus and a new species of betasatellite infecting Carica papaya in India. The sequences of the begomovirus and betasatellite show 90.03% nucleotide sequence identity to an isolate of Radish leaf curl virus and 92.25% identity to an isolate of Tomato leaf curl betasatellite, respectively. Maximum likelihood phylogenetic tree of begomovirus sequence of isolate DP2 (KX353622) showed distant relationships with previously characterised begomoviruses. Recombination analysis proposed six recombination breakpoints in begomovirus genome with other geographical begomovirus isolates.     

固定化酵母发酵木瓜-西番莲复合果酒工艺的研究

[目的]研究固定化酵母发酵木瓜-西番莲复合果酒的最佳工艺条件。[方法]以木瓜和西番莲果为原料,研究了初始加糖量、pH、固定化酵母接种量、发酵温度对木瓜-西番莲复合果酒残糖、酒精度和感官品质的影响。[结果]影响木瓜西番莲复合果酒发酵残糖、酒精度和感官品质的主要因素为复合果汁初始加糖量,最佳发酵条件为混合果汁起始糖度28%、pH 4.5、酵母接种量0.015%、发酵温度25℃。[结论]在最佳工艺条件下可获得色泽淡黄自然、拥有着和谐的果香与酒香、入口清爽、酒精含量约13.8%的木瓜-西番莲复合果酒。     

农杆菌浸润瞬时表达分析番木瓜miR162a的研究

农杆菌浸润瞬时表达方法不仅在蛋白互作、鉴定基因沉默抑制子及基因功能研究方面得到广泛应用,目前还应用在小RNA的研究中.在对植物microRNA的研究中,农杆菌浸润方法通过将将转基因导入农杆菌浸润本生烟瞬时表达,可以快速、稳定地对植物microRNA进行检测和鉴定,样品在几天内就可分析完成,比获得稳定遗传的转基因有优势.笔者借鉴国外拟南芥microRNA瞬时表达分析的方法,克隆了番木瓜的miR62a前体序列连接到双元表达载体pSuper1300上构建成35S::cpa-miR162a质粒,导人农杆菌GV3101后浸润本生烟,浸润后48 h提取本生烟叶片总RNA后通过miRNA northern blot检测到miR162a,而本生烟和番木瓜自身则无法检测到低丰度的miR162a.本文介绍的农杆菌浸润瞬时表达方法能够在体外加工产生成熟的miRNA,该技术有助于对microRNA的结构-功能的相关性及靶标验证进一步研究.     

Sap flow in papaya plants: Laboratory calibrations and relationships with gas exchanges under field conditions

In papaya plants a study to quantify the water flow through the trunk is important for to promote a good water management in commercial orchard. The objective was to study the relationship between water flow through the trunk and temperature measurements determined by probes inserted in the papaya plant stem in laboratory. In addition, was possible to study the relationship between sap flow and instantaneous gas exchange in field conditions. We constructed an instrument that maintained a stable water flux through a 0.30 m stem section with a constant pressure, simulating the xylem sap flow through the stem. Water flux was adjusted by varying pressure of water in the stem section. The mathematical model used to fit the relation between K (Granier heat coefficient) values and sap flow density was the exponential model: u = 0.5511 × K^1.9104. Field studies was conducted in a commercial orchard located in North of the State of Rio de Janeiro, with 12 plants in October 2002, and eight plants in January 2003. We verified that instantaneous transpiration, measured by a portable system of gas exchange (porometry), presented a good (R² = 0.75) positive relationship with xylem sap flow. Estimates of papaya sap flow can be obtained by scaling portable photosynthesis system measurements with exposed leaves, however the relationship is non-linear in higher instantaneous transpiration rates. The causes of the non-linear relationship in higher transpiration are discussed. In addition, was possible to obtain a good (R² = 0.76) relationship between net photosynthesis rate and xylem sap flow in papaya field-grown.