Echemendía A L, Ramos P L, Peral R, Fuentes A, González G, Sanpedro J, Morales F
First and fifth authors, Instituto de Investigaciones de Sanidad Vegetal, Cuba.
second, third, and fourth authors, Centro de Ingeniería Genética y Biotecnología, P.O. Box 6162. La Habana. CP 10600. Cuba. E-mail:
Plant Dis. 2001 Sep;85(9):1030. doi: 10.1094/PDIS.2001.85.9.1030C.
In Cuba, the emergence of bean golden mosaic was associated with high populations of Bemisia tabaci in common bean (Phaseolus vulgaris L.) plantings in the 1970s (1). During the last two decades, the disease has caused significant economic losses, forcing some growers to abandon bean production. In Holguín, one of the main bean producing provinces of the country, about 2,000 ha of beans were abandoned in 1991 due to the high incidence of this whitefly-transmitted virus. At that time, yield losses associated with this disease reached 90 to 100% in farmer's fields. In spite of various control measures, the disease affected 33, 28, and 6.5% of the total area planted in Cuba to common bean in 1990, 1992, and 1996, respectively. For this investigation, common bean leaves showing systemic yellowing symptoms were collected in fields located in the provinces of Havana, Matanzas, and Holguín during 1998-1999. Sap and total DNA leaf extracts were used to inoculate healthy bean plants by manual and biolistic procedures, respectively. Characteristic yellowing symptoms were more efficiently reproduced using a particle gun device than by manual inoculation (18/20 plants and 5/20 plants, respectively, for a Holguín virus isolate). DNA extracts were further analyzed by polymerase chain reaction using two degenerate primer sets: PAL1v1978-PAR1c715 and PAL1c1960-PAR1v722 (2). Fragments of approximately 1.4 and 1.2 kb were amplified and cloned. Restriction fragment length polymorphism analysis of the cloned 1.4-kb fragments was performed with BglII, HincII, SalI, EcoRI, PstI, and XbaI, indicating that selected isolates from the three Cuban provinces shared identical restriction patterns. The nucleotide sequence obtained from two clones of a virus isolate from Holguín, was compared to sequences available for other begomoviruses using BLAST. The Cuban isolate shared up to 94% nt sequence identity with various strains of Bean golden yellow mosaic virus (BGYMV) in the first 250 nt of the rep gene. For the common region (CR), scores were 93% for BGYMV-GA (Guatemala), 92% for BGYMV-MX (southern Mexico) and BGYMV-PR (Puerto Rico), and 91% for BGYMV-DR (Dominican Republic). The iterative sequence ATGGAG was identified in the CR of the Cuban BGYMV isolate, as reported for other BGYMV isolates. Finally, the Cuban begomovirus, hereafter referred to as BGYMV-CU, shared nt and aa sequence identities of 94 and 100%, respectively, with the coat protein gene of BGYMV-MX. We conclude that the begomovirus isolated from mosaic-affected common bean plants in the province of Holguín is a member of the Mesoamerican BGYMV group (3). References: (1) N. Blanco and C. Bencomo. Cienc. Agric. 2:39, 1978. (2) M. R. Rojas et al. Plant Dis. 77:340, 1993. (3) Morales and Anderson, Arch. Virol. 146:415, 2001.
在古巴,菜豆金色花叶病的出现与20世纪70年代普通菜豆(Phaseolus vulgaris L.)种植中烟粉虱的高虫口密度有关(1)。在过去的二十年里,这种病害造成了重大经济损失,迫使一些种植者放弃菜豆生产。在该国主要菜豆生产省份之一的奥尔金省,1991年约2000公顷的菜豆因这种粉虱传播病毒的高发病率而被弃种。当时,在农民的田地里,与这种病害相关的产量损失达到了90%至100%。尽管采取了各种防治措施,但在1990年、1992年和1996年,这种病害分别影响了古巴普通菜豆种植总面积的33%、28%和6.5%。在1998 - 1999年期间,从哈瓦那省、马坦萨斯省和奥尔金省的田间收集了表现出系统黄化症状的普通菜豆叶片。汁液和叶片总DNA提取物分别通过手工和基因枪接种程序用于接种健康菜豆植株。使用基因枪装置比手工接种更有效地再现了特征性黄化症状(对于一种奥尔金病毒分离株,分别为18/20株和5/20株)。DNA提取物通过聚合酶链反应使用两组简并引物进一步分析:PAL1v1978 - PAR1c715和PAL1c1960 - PAR1v722(2)。扩增并克隆了约1.4 kb和1.2 kb的片段。用BglII、HincII、SalI、EcoRI、PstI和XbaI对克隆的1.4 kb片段进行限制性片段长度多态性分析,表明从古巴三个省份选择的分离株具有相同的限制性图谱。使用BLAST将从奥尔金省一个病毒分离株的两个克隆获得的核苷酸序列与其他双生病毒的可用序列进行比较。古巴分离株在rep基因的前250个核苷酸中与菜豆金黄花叶病毒(BGYMV)的各种菌株共享高达94%的核苷酸序列同一性。对于共同区域(CR),与BGYMV - GA(危地马拉)的得分是93%,与BGYMV - MX(墨西哥南部)和BGYMV - PR(波多黎各)的得分是92%,与BGYMV - DR(多米尼加共和国)的得分是91%。如其他BGYMV分离株报道的那样,在古巴BGYMV分离株的CR中鉴定出了重复序列ATGGAG。最后,古巴双生病毒,以下简称BGYMV - CU,与BGYMV - MX的外壳蛋白基因的核苷酸和氨基酸序列同一性分别为94%和100%。我们得出结论,从奥尔金省受花叶病影响的普通菜豆植株中分离出的双生病毒是中美洲BGYMV组的成员(3)。参考文献:(1)N. Blanco和C. Bencomo。Cienc. Agric. 2:39,1978。(2)M. R. Rojas等人。Plant Dis. 77:340,1993。(3)Morales和Anderson,Arch. Virol. 146:415,2001。