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First Report of Sooty Blotch and Flyspeck Caused by Species of Dissoconium, MycosphaereUa, and Peltaster on Hawthorn Fruit in China

机译:中国山楂果实中的二毒,鼠疫菌和小白菜引起的煤烟斑和蝇斑的首次报道

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Sooty blotch and flyspeck (SBFS), a disease complex comprised of as many as 30 putative species of fungi, occurs on the cuticle of pome fruits in moist production regions worldwide, inciting cosmetic damage that causes significant economic losses. Chinese hawthorn (Crataegus pinnatifida Bge.) is an economically important tree species in China. Its fruit are sold fresh or dried and are used as a culinary spice as well as an ingredient in Chinese traditional medicine. In October of 2007, Chinese hawthorn fruit exhibiting SBFS signs were sampled from supermarkets in Yangling, Shaanxi Province and Luoyang, Henan Province, China. Thalli directly from the hawthorn fruit were transferred onto potato dextrose agar (PDA) slants under a dissecting microscope and cultured at 22 plus or minus 1 degree C in darkness. DNA was extracted from pure isolates and the internal tran-scribed spacer (ITS) region of the nuclear ribosomal DNA (nrDNA) was amplified and sequenced using primers ITS-1F and ITS4. Phylogenetic analysis of the ITS sequences revealed that the 35 isolates generated in this study included five species in three genera: Dissoconium sp. (18 isolates), Mycosphaerella sp. (5 isolates), and Peltaster sp. 1 (4 isolates), Peltaster sp. 2 (4 isolates), and Peltaster sp. 3 (4 isolates). To fulfill Koch's postulates and verify that these fungi could also infest apple fruit, two representative isolates of each putative species were inoculated onto mature intact hawthorn and apple (cv. Fuji) fruit that had been surface disinfested with 75% ethanol and allowed to dry. Inoculum was prepared by comminuting 1-month-old cultures growing on PDA into a suspension of mycelial fragments and conidia (10 super(5) to similar to 10 super(6) CFU/ml) in a blender with sterile deionized water (SDW). Each isolate was inoculated on three hawthorn and three apple fruit by using cotton swabs. As controls, two surface-disinfested hawthorn and apple fruit were swabbed with SDW. After the inoculated hawthorn and apple fruit had been incubated in a moist chamber at 22 plus or minus 1 degree C for 1 month, all inoculated hawthorn and apple fruit exhibited SBFS signs similar to those of the original colonies on hawthorn fruit, but the controls did not. Reservoir hosts have been inferred to play an important role in SBFS by providing the fungi with overwintering habitat and inoculum for infestations on apple. Many reservoir hosts have been reported in the United States and Japan. To our knowledge, this is the first report of fungi in the SBFS complex on hawthorn fruit and the first confirmation that fungi growing on hawthorn fruit can produce SBF5 signs on apple fruit. These results identify hawthorn as a potential inoculum source for SBFS in apple orchards.
机译:乌贼斑点和蝇斑(SBFS)是一种由多达30种推定真菌组成的疾病复合物,在世界各地潮湿的生产地区的梨果的表皮上发生,造成化妆品损害,造成重大的经济损失。中国山楂(Crataegus pinnatifida Bge。)是中国经济上重要的树种。它的果实可以新鲜或干燥出售,可以用作烹饪香料以及中药成分。 2007年10月,从陕西省杨凌市和河南省洛阳市的一家超市中取样了带有SBFS标志的山楂果。在解剖显微镜下,将来自山楂果实的Thalli直接转移到马铃薯葡萄糖琼脂(PDA)斜面上,并在黑暗中于22摄氏度或-1摄氏度下培养。从纯分离株中提取DNA,并使用引物ITS-1F和ITS4扩增核糖体DNA(nrDNA)的内部转录间隔区(ITS)。对ITS序列进行的系统发育分析表明,在这项研究中产生的35个分离株包括三个属中的5个种:Dissoconium sp。 (18个分离株),Mycosphaerella sp。 (5个分离株)和Peltaster sp。 1(4株),Peltaster sp。 2(4个分离株)和Peltaster sp。 3(4个隔离株)。为了满足Koch的假设并验证这些真菌也可能侵染苹果果实,将每种推定物种的两个代表性分离株接种到成熟的完整山楂和苹果果实上,该果实在表面用75%乙醇除虫并干燥。通过在无菌无菌去离子水(SDW)的搅拌器中将在PDA上生长的1个月大的培养物粉碎成菌丝体碎片和分生孢子(10 super(5)类似于10 super(6)CFU / ml)的悬浮液来制备接种物。 。使用棉签将每个分离株接种在三个山楂和三个苹果果实上。作为对照,用SDW擦拭了两个表面消毒过的山楂和苹果果实。将接种过的山楂和苹果果实在22摄氏度和-1摄氏度的潮湿室内孵育1个月后,所有接种过的山楂和苹果果实均显示出与山楂果实上原始菌落相似的SBFS征兆,但对照组确实做到了不。通过为真菌提供越冬的生境和接种物来侵染苹果,已经推断出水库宿主在SBFS中起重要作用。在美国和日本,已经报道了许多储层宿主。据我们所知,这是山楂果实SBFS复合物中的真菌的首次报道,也是山楂果实上生长的真菌可在苹果果实上产生SBF5迹象的第一个证实。这些结果表明山楂是苹果园中SBFS的潜在接种源。

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