三裂葉豚草霜霉菌（Plasmopara angustiterminalis）鑒定及其與銹菌（Puccinia xanth
發(fā)布時(shí)間：2018-08-24 12:58
【摘要】：三裂葉豚草(Ambrosia trifida Linn.)原產(chǎn)于北美美國(guó)和墨西哥交界Sonoland地區(qū),現(xiàn)已傳至歐洲、亞洲、澳洲等許多國(guó)家和地區(qū)。三裂葉豚草可嚴(yán)重影響農(nóng)作物生長(zhǎng)和農(nóng)作物產(chǎn)量,并嚴(yán)重危害人類身體健康,是世界性外來(lái)入侵有害生物,已給各國(guó)造成了嚴(yán)重的生態(tài)安全威脅。我國(guó)是三裂葉豚草等外來(lái)入侵有害生物危害最為嚴(yán)重的國(guó)家之一,尤其以遼寧為主要發(fā)生危害地的東北地區(qū)受其影響最為嚴(yán)重。本文通過(guò)光學(xué)顯微鏡、掃描電子顯微鏡和透射電子顯微鏡,研究銹菌和霜霜菌侵染后的三裂葉豚草葉片,發(fā)現(xiàn)兩種蒼耳柄銹菌三裂葉豚草�；�(Puccinia xanthii.f.sp.ambrosiae-trifidae S.W.T.Batra)菌絲和蒼耳軸霜霉(Plasmopara agustiterminalis Novotelnova)菌絲交錯(cuò)寄生在下表皮相連的葉肉海綿組織細(xì)胞內(nèi);同時(shí)發(fā)現(xiàn)霜霉菌菌絲在葉肉細(xì)胞內(nèi)特化形成吸器并進(jìn)入相鄰葉肉細(xì)胞內(nèi)吸取養(yǎng)分;未觀察到生長(zhǎng)在一起的兩種真菌菌絲或者吸器結(jié)構(gòu)有相互作用。為了進(jìn)一步揭示共同侵染三裂葉豚草的蒼耳柄銹菌三裂葉豚草�；秃蜕n耳軸霜霉是否存在相互侵染,利用熒光原位雜交技術(shù),針對(duì)蒼耳柄銹菌三裂葉豚草�；秃蜕n耳軸霜霉專性寄生菌設(shè)計(jì)寡核苷酸探針和輔助探針進(jìn)行雙標(biāo)記,以石蠟切片為介導(dǎo),對(duì)雜交探針濃度及雜交試驗(yàn)條件進(jìn)行了摸索和優(yōu)化。石蠟切片雜交預(yù)處理各步驟時(shí)間由5分鐘縮短為3 min,雜交探針濃度1 μg/mL、2 μg/mL、4 μg/mL,蛋白酶K處理5 min、10 min、15 min;幾丁質(zhì)酶酶解 10 min、30 min、60 min,DAPI 復(fù)染濃度分別為 1 μg/mL、4 μg/mL,結(jié)果表明:在探針濃度為2μg/mL、蛋白酶K36℃處理10min、幾丁質(zhì)酶36℃酶解30 min、DAPI復(fù)染濃度4 μg/mL時(shí)試驗(yàn)結(jié)果顯示霜霉菌菌絲穿插寄生在植物葉片中,銹菌夏孢子基部菌絲集中分布于接近下表皮葉肉海綿組織細(xì)胞,霜霉菌孢子囊及孢子梗均可以顯示熒光。
[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，
本文編號(hào)：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，
本文編號(hào)：2200889
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