Duan Y X, Wang D, Chen L J
Nematology Institute of Northern China, College Plant Protection, Shenyang Agricultural University, Shenyang 110866, China.
Plant Dis. 2012 Sep;96(9):1385. doi: 10.1094/PDIS-04-12-0374-PDN.
During a survey for cyst nematodes from August to September 2011, nearly 15% of cactus (Cereus jamacaru) plants exhibited yellowing and wilting symptoms in greenhouses from Shenyang Botanical Garden, Liaoning Province, China. Cysts (averaging 50 per 100 g of samples) were detected by the sieving-decanting method on the roots and in rhizospheric soil. Second stage juveniles and eggs were isolated directly from cysts. Cysts, juveniles, and eggs were identified by morphology. Cysts (n = 12) were rounded to lemon-shaped with a protruding neck and vulva. The cyst wall had a zig-zag pattern. The vulval cone was circumfenestrate without underbridge and bullae but generally with vulval denticles. The cysts were characterized by body length excluding neck (range = 399.5 to 622.0 μm, mean = 524.9), body width (300.4 to 469.9 μm, 383.4), length to width ratio (1.1 to 1.7, 1.4), neck length (41.0 to 130.9 μm, 61.5) and width (55.0 to 98.7 μm, 76), and circumfenestral diameter length (24.6 to 30.2 μm, 28.4). Measurements of second-stage juveniles (n = 20) included length of body (range = 467.3 to 542.5 μm, mean = 513.8), stylet (23.0 to 25.8 μm, 24.6) with knobs rounded to slightly projecting anteriorly and concave on anterior surface, tail (45.9 to 59.5 μm, 52.2), and hyaline tail terminal (16.3 to 23.2 μm, 19.1). Eggs (n = 20) had heavy punctations on the shell surface. All morphological data and characteristics were consistent with Cactodera cacti (3). Molecular evidence confirmed the identification. DNA from a single cyst was extracted by using the protocol described by Subbotin et al. (2), the rDNA-internal transcribed spacer (ITS) and D2-D3 fragments of the 28S rDNA were amplified with universal primers TW81 (5'-GTTTCCGTAGGTGAACCTGC-3') and AB28 (5'-ATATGCTTAAGTTCAGCGGGT-3'), D2A(5'-ACAAGTACCGTGAGGGAAAGTTG-3') and D3B(5'-TCGGAAGGAACCAGCTACTA-3'), respectively. The ITS PCR product was digested with eight restriction enzymes (AluI, AvaI, Bsh1236I, BsuRI, CfoI, MvaI, PstI, and RsaI) to obtain restriction fragment length polymorphism profiles (4). The ITS and D2D3 sequences were cloned and assayed using an ABI-PRISM 3730 Genetic Analyzer (Applied Sangon, Shanghai, China) and were subjected to a database search using BLAST (National Centre for Biotechnology Information). The 980-bp ITS sequence exhibited 99% similarity with that of a C. cacti isolate from Iran (GenBank Accession No. AF498393.1) and the 787-bp D2D3 sequence exhibited 99% similarity with a C. cacti isolate from Germany (GenBank Accession No. DQ328702.1). Cactus cyst nematode has been mainly reported on ornamental plants of the family Cactaceae grown in greenhouses. Infested plants become reddish brown to yellow in color, wilted, stunted, with reduced flower production and shortening flower period. With high population densities of C. cacti, death of plants may occur (1). To the best of our knowledge, this is the first report of C. cacti in northern China. References: (1) R. P. Esser. Division of Plant Industry, 197:3, 1992. (2) S. A. Subbotin et al. Nematology, 2:153, 2000. (3) S. A. Subbotin et al. Systematics of Cyst Nematodes (Nematoda: Heteroderinae). Volume 8 Part A. Brill, Leiden, the Netherlands, 2010. (4) M. Z. Tanha et al. Nematology, 5:99, 2003.
2011年8月至9月对孢囊线虫进行调查期间,在中国辽宁省沈阳植物园的温室中,近15%的仙人掌(Cereus jamacaru)植株出现黄化和萎蔫症状。采用过筛倾析法在根部和根际土壤中检测到孢囊(每100克样品平均有50个)。直接从孢囊中分离出第二阶段幼虫和卵。通过形态学对孢囊、幼虫和卵进行鉴定。孢囊(n = 12)呈圆形至柠檬形,颈部和阴门突出。孢囊壁呈锯齿状。阴门锥有周侧孔,无下桥和疱,但通常有阴门小齿。孢囊的特征为不包括颈部的体长(范围= 399.5至622.0μm,平均= 524.9)、体宽(300.4至469.9μm,383.4)、长宽比(1.1至1.7,1.4)、颈部长度(41.0至130.9μm,61.5)和宽度(55.0至98.7μm,76)以及周侧孔直径长度(24.6至30.2μm,28.4)。第二阶段幼虫(n = 20)的测量数据包括体长(范围= 467.3至542.5μm,平均= 513.8)、口针(23.0至25.8μm,24.6),其瘤呈圆形至稍向前突出且前表面凹陷,尾部(45.9至59.5μm,52.2)以及透明尾端(16.3至23.2μm,19.1)。卵(n = 20)在壳表面有密集的小孔。所有形态学数据和特征均与仙人掌孢囊线虫(Cactodera cacti)一致(3)。分子证据证实了该鉴定结果。使用Subbotin等人(2)描述的方法从单个孢囊中提取DNA,分别用通用引物TW81(5'-GTTTCCGTAGGTGAACCTGC-3')和AB28(5'-ATATGCTTAAGTTCAGCGGGT-3')、D2A(5'-ACAAGTACCGTGAGGGAAAGTTG-3')和D3B(5'-TCGGAAGGAACCAGCTACTA-3')扩增28S rDNA的rDNA内部转录间隔区(ITS)和D2 - D3片段。ITS PCR产物用8种限制性内切酶(AluI、AvaI、Bsh1236I、BsuRI、CfoI、MvaI、PstI和RsaI)消化以获得限制性片段长度多态性图谱(4)。将ITS和D2D3序列克隆并使用ABI - PRISM 3730遗传分析仪(Applied Sangon,上海,中国)进行检测,并使用BLAST(美国国立生物技术信息中心)在数据库中进行搜索。980 bp的ITS序列与来自伊朗的仙人掌孢囊线虫分离株(GenBank登录号AF498393.1)的序列相似性为99%,787 bp的D2D3序列与来自德国的仙人掌孢囊线虫分离株(GenBank登录号DQ3A28702.1)的序列相似性为99%。仙人掌孢囊线虫主要报道于温室中种植的仙人掌科观赏植物上。受侵染的植物会变成红棕色至黄色,萎蔫、发育不良,花朵产量降低且花期缩短。当仙人掌孢囊线虫种群密度较高时,植物可能会死亡(1)。据我们所知,这是中国北方首次报道仙人掌孢囊线虫。参考文献:(1)R. P. Esser。植物产业司,197:X3,1992。(2)S. A. Subbotin等人。线虫学,2:153,2000。(3)S. A. Subbotin等人。孢囊线虫的系统学(线虫纲:异皮科)。第8卷A部分。Brill,荷兰莱顿,2010。(4)M. Z. Tanha等人。线虫学,5:99,2003。