三裂叶豚草霜霉菌（Plasmopara angustiterminalis）鉴定及其与锈菌（Puccinia xanth
发布时间：2018-08-24 12:58
【摘要】：三裂叶豚草(Ambrosia trifida Linn.)原产于北美美国和墨西哥交界Sonoland地区,现已传至欧洲、亚洲、澳洲等许多国家和地区。三裂叶豚草可严重影响农作物生长和农作物产量,并严重危害人类身体健康,是世界性外来入侵有害生物,已给各国造成了严重的生态安全威胁。我国是三裂叶豚草等外来入侵有害生物危害最为严重的国家之一,尤其以辽宁为主要发生危害地的东北地区受其影响最为严重。本文通过光学显微镜、扫描电子显微镜和透射电子显微镜,研究锈菌和霜霜菌侵染后的三裂叶豚草叶片,发现两种苍耳柄锈菌三裂叶豚草专化型(Puccinia xanthii.f.sp.ambrosiae-trifidae S.W.T.Batra)菌丝和苍耳轴霜霉(Plasmopara agustiterminalis Novotelnova)菌丝交错寄生在下表皮相连的叶肉海绵组织细胞内;同时发现霜霉菌菌丝在叶肉细胞内特化形成吸器并进入相邻叶肉细胞内吸取养分;未观察到生长在一起的两种真菌菌丝或者吸器结构有相互作用。为了进一步揭示共同侵染三裂叶豚草的苍耳柄锈菌三裂叶豚草专化型和苍耳轴霜霉是否存在相互侵染,利用荧光原位杂交技术,针对苍耳柄锈菌三裂叶豚草专化型和苍耳轴霜霉专性寄生菌设计寡核苷酸探针和辅助探针进行双标记,以石蜡切片为介导,对杂交探针浓度及杂交试验条件进行了摸索和优化。石蜡切片杂交预处理各步骤时间由5分钟缩短为3 min,杂交探针浓度1 μg/mL、2 μg/mL、4 μg/mL,蛋白酶K处理5 min、10 min、15 min;几丁质酶酶解 10 min、30 min、60 min,DAPI 复染浓度分别为 1 μg/mL、4 μg/mL,结果表明:在探针浓度为2μg/mL、蛋白酶K36℃处理10min、几丁质酶36℃酶解30 min、DAPI复染浓度4 μg/mL时试验结果显示霜霉菌菌丝穿插寄生在植物叶片中,锈菌夏孢子基部菌丝集中分布于接近下表皮叶肉海绵组织细胞,霜霉菌孢子囊及孢子梗均可以显示荧光。
[Abstract]:Ragweed (Ambrosia trifida Linn.) Native to North America, the United States and Mexico on the border of Sonoland, has been spread to Europe, Asia, Australia and many other countries and regions. Ragweed (ragweed trifida) can seriously affect crop growth and crop yield and seriously endanger human health. It is an alien invasive pest in the world and has caused a serious threat to the ecological security of various countries. China is one of the countries where invasive pests such as ragweed artemisia trifida are most seriously damaged, especially in Northeast China, where Liaoning is the main place of damage. By means of optical microscope, scanning electron microscope and transmission electron microscope, the leaf of ragweed trifida after infection with Ruccinia trifida was studied. It was found that two species of Ruccinia trifida (Puccinia xanthii.f.sp.ambrosiae-trifidae S.W.T.Batra) mycelium and (Plasmopara agustiterminalis Novotelnova) mycelium intersected in the mesophyll sponge cells connected to the lower epidermis. At the same time, it was found that the hyphae inosculated the mesophyll cells to form an absorber and to absorb nutrients into the adjacent mesophyll cells, and no interaction between the hyphae or the haustorium structure of the two fungi growing together was observed. In order to further reveal the mutual infection of Ruccinia trifida and Rhizoma artemisia trifida, the fluorescence in situ hybridization (fish) technique was used. Oligonucleotide probes and auxiliary probes were designed to double label the specific parasites of ragweed trifida and Pleurotus sibiricus. The concentration of hybridization probe and the conditions of hybridization test were explored and optimized with paraffin sections as the mediators. The pretreatment time of paraffin section hybridization was shortened from 5 minutes to 3 min, hybridization probe concentration of 1 渭 g / mL ~ 2 渭 g / mL ~ (2 渭 g 路m ~ (-1) and 4 渭 g / mL of protease K treatment for 5 min,10 min,15 min; chitinase hydrolysis for 10 min,30 min,60 min,DAPI respectively. The results showed that: 1 渭 g 路mL ~ (-1) 路min ~ (-1) 路min ~ (-1), respectively. When the probe concentration was 2 渭 g / mL, protease K36 鈩，
本文编号：2200889
[Abstract]:Ragweed (Ambrosia trifida Linn.) Native to North America, the United States and Mexico on the border of Sonoland, has been spread to Europe, Asia, Australia and many other countries and regions. Ragweed (ragweed trifida) can seriously affect crop growth and crop yield and seriously endanger human health. It is an alien invasive pest in the world and has caused a serious threat to the ecological security of various countries. China is one of the countries where invasive pests such as ragweed artemisia trifida are most seriously damaged, especially in Northeast China, where Liaoning is the main place of damage. By means of optical microscope, scanning electron microscope and transmission electron microscope, the leaf of ragweed trifida after infection with Ruccinia trifida was studied. It was found that two species of Ruccinia trifida (Puccinia xanthii.f.sp.ambrosiae-trifidae S.W.T.Batra) mycelium and (Plasmopara agustiterminalis Novotelnova) mycelium intersected in the mesophyll sponge cells connected to the lower epidermis. At the same time, it was found that the hyphae inosculated the mesophyll cells to form an absorber and to absorb nutrients into the adjacent mesophyll cells, and no interaction between the hyphae or the haustorium structure of the two fungi growing together was observed. In order to further reveal the mutual infection of Ruccinia trifida and Rhizoma artemisia trifida, the fluorescence in situ hybridization (fish) technique was used. Oligonucleotide probes and auxiliary probes were designed to double label the specific parasites of ragweed trifida and Pleurotus sibiricus. The concentration of hybridization probe and the conditions of hybridization test were explored and optimized with paraffin sections as the mediators. The pretreatment time of paraffin section hybridization was shortened from 5 minutes to 3 min, hybridization probe concentration of 1 渭 g / mL ~ 2 渭 g / mL ~ (2 渭 g 路m ~ (-1) and 4 渭 g / mL of protease K treatment for 5 min,10 min,15 min; chitinase hydrolysis for 10 min,30 min,60 min,DAPI respectively. The results showed that: 1 渭 g 路mL ~ (-1) 路min ~ (-1) 路min ~ (-1), respectively. When the probe concentration was 2 渭 g / mL, protease K36 鈩，
本文编号：2200889
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