本文選題：天祝草原 + 禾谷孢囊線蟲��； 參考：《甘肅農(nóng)業(yè)大學》2015年碩士論文

【摘要】：禾谷孢囊線蟲(Cereal cyst nematode,CCN Heterodera SPP.)是一類危害小麥(Triticum aestivum)、大麥(Hordeum vulgare)、燕麥(Avena sativa)等禾谷類作物的世界性重要病原線蟲。禾谷孢囊線蟲除為害小麥、大麥和燕麥等禾谷類作物外,一些多年生的禾本科植物科也可作為禾谷孢囊線蟲的中間寄主或橋梁寄主。本研究根據(jù)甘肅省天祝草原農(nóng)牧區(qū)的分布特征,系統(tǒng)的對禾谷孢囊線蟲在天祝高寒草甸草原的發(fā)生、生態(tài)分布、孢囊線蟲種類及優(yōu)勢種群進行了研究。2013-2014年,采用隨機采樣法,對甘肅省天祝高寒草甸草原主要牧區(qū)的禾谷孢囊線蟲進行了較系統(tǒng)的調(diào)查,調(diào)查范圍覆蓋了天�？h主要牧區(qū)和半農(nóng)半牧區(qū)的13個鄉(xiāng)鎮(zhèn)。采集樣共計本248份,其中56份樣品中檢出有孢囊線蟲,占22.6%。通過鏡檢,所調(diào)查的13個鄉(xiāng)鎮(zhèn),7個鄉(xiāng)鎮(zhèn)土樣有孢囊線蟲檢出,檢出率由高到低分別為打柴溝鎮(zhèn)47.4%、塞拉隆鄉(xiāng)46.7%、抓喜秀龍鄉(xiāng)44.9%、石門鎮(zhèn)40.0%、安遠鎮(zhèn)34.4%、大紅溝鄉(xiāng)11.1%和旦馬鄉(xiāng)9.8%。炭山嶺鎮(zhèn)、塞什斯鎮(zhèn)、天堂鎮(zhèn)、松山鎮(zhèn)、祁連鄉(xiāng)和哈溪鎮(zhèn)土樣中孢囊線蟲檢出率為0。抓喜秀龍鄉(xiāng)一采樣地點孢囊基數(shù)最高為54個/100 g土樣。通過以0?10 cm、10?20 cm、20?30 cm、30?40 cm為垂直采樣標準,在已發(fā)現(xiàn)有孢囊線蟲的40個樣點進行分層采樣研究天祝草原孢囊線蟲的垂直分布情況,結(jié)果表明,孢囊線蟲均布在0?10 cm和10?20 cm土層中,30 cm以下土樣中未檢出孢囊線蟲,且所有土樣中0?10 cm孢囊線蟲密度均大于對應(yīng)土樣的10?20 cm中孢囊密度。本研究應(yīng)用形態(tài)學結(jié)合分子生物學鑒定,鑒定結(jié)果表明,甘肅天祝高寒草甸草原孢囊線蟲優(yōu)勢種群為禾谷孢囊線蟲(Heteroder avenae)。其形態(tài)學特征為孢囊淺褐色,闊檸檬形,陰門錐雙膜孔,未見陰門下橋,泡狀突明顯。利用通用引物AB28和TW81,通過PCR技術(shù)對禾谷孢囊線蟲群體的核糖體基因的內(nèi)轉(zhuǎn)錄間隔區(qū)(rDNA-ITS)序列的擴增,后經(jīng)PCR產(chǎn)物純化回收、克隆、測序,得到ITS區(qū)序列長度為1045 bp,序列中無堿基插入和缺失,均有堿基替換。將所測得序列在Genbank和Blast上比對,下載近緣種序列,在MEGA5.2中運用UPGMA法構(gòu)建系統(tǒng)發(fā)育樹,結(jié)果顯示,此禾谷孢囊線蟲群體與H.avena群體相似度達99%~100%,進一步證實寄生于甘肅天祝高寒草甸草原牧草孢囊線蟲均為H.avenae。ITS構(gòu)建的系統(tǒng)發(fā)育樹表明,本研究線蟲群體與中國的禾谷孢囊線蟲、俄羅斯的早熟禾孢囊線蟲(H.pratensis)和德國的早熟禾孢囊線蟲(H.pratensis)群體同時聚為一個小的分支,揭示了它們之間具有較高的親緣關(guān)系。為了測定天祝牧草禾谷孢囊線蟲(H.avenae)群體對小麥的致病力,采用室內(nèi)人工接種的方法,將分離自嵩草和矮嵩草的2個禾谷孢囊線蟲孢囊經(jīng)低溫處理后,將線蟲卵接種至小麥“矮抗58”,每株3000粒卵,將接種的小麥苗置于15±2℃的培養(yǎng)室中,用鏑燈燈管補光,每天光照14 h。1周后,溫度調(diào)至18±2℃。定期澆水,使PVC管內(nèi)的土壤保持一定的濕度,75天后觀察統(tǒng)計小麥根部和根際土壤孢囊數(shù)。結(jié)果表明,天祝牧草禾谷孢囊線蟲群體可以侵染小麥。
[Abstract]:Cereal cyst nematode (CCN Heterodera SPP.) is a class of worldwide important pathogenic nematodes that are harmful to wheat (Triticum aestivum), barley (Hordeum vulgare), oat (Avena sativa) and other cereal crops. The cereal cyst nematode is a perennial grasses except for wheat, barley, and oats. This study, based on the distribution characteristics of the farm and pastoral areas of Tianzhu grassland in Gansu Province, studied the occurrence, ecological distribution, the species of cyst nematode and the dominant population of the cyst nematode in Tianzhu meadow grassland in Gansu province. The random sampling method was applied to Gansu. A systematic investigation was conducted in the main pastoral areas of the main pasture pasture of Tianzhu alpine meadow, which covered 13 villages and towns in the main pastoral areas and half agricultural and semi pastoral areas of Tianzhu County. There were 248 samples collected in the main pastoral areas of Tianzhu County. Among them, 56 samples were found with cyst nematode, accounting for 22.6%. through microscopic examination, 13 villages and towns and 7 villages and towns. The detection rate of cyst nematode from high to low were 47.4% in Chai Gou Town, 46.7% in saailon Township, 44.9% in Xi Xiu long Township, 40% in Shimen Town, 34.4% in Anyuan Town, 11.1% in Dagu Township, 9.8%. in denaten Township, sash Town, Paradise Town, Songshan Town, Qilian Township and Haxi Town, 0. The highest cysts were 54 /100 g soil samples. By using 0? 10 cm, 10? 20 cm, 20? 30 cm, 30? 40 cm as vertical sampling criteria, the vertical distribution of the cyst nematode in Tianzhu steppe was studied by stratified sampling at 40 samples of the cyst nematode. The results showed that the cyst nematode was distributed in the 0? 10 cm and 10? 20 cm soil. No cyst nematode was detected, and the density of 0? 10 cm cyst nematode in all soil samples was greater than that of 10? 20 cm in the corresponding soil samples. The results showed that the dominant population of Gansu Tianzhu alpine meadow steppe was that the dominant population of the cyst nematode in the meadow was Heteroder avenae. The cysts were light brown, broad lemonade and double membrane holes in the cones of the vulva. There were no inferior pons and obvious alveolar processes. Using universal primers AB28 and TW81, the PCR technique was used to amplify the internal transcriptional interval (rDNA-ITS) sequence of the ribosome gene of the cyst nematode population, and then the PCR product was purified and sequenced, and the sequence length of the ITS region was 1045 BP, The sequences were inserted and missing with base substitutions. The sequences measured on Genbank and Blast were compared, the sequences were downloaded and the phylogenetic tree was constructed by UPGMA method in MEGA5.2. The results showed that the similarity of this colony of Cereal Cyst Nematode with H.avena population was 99%~100%, further confirmed parasitic in Gansu Tianzhu alpine meadow. The phylogenetic tree of the original herbage bursa nematode is H.avenae.ITS, which shows that the nematode population of this study conforms to a small branch of China's cereal cyst nematode, Russian precocious bursa nematode (H.pratensis) and German precocious bursa nematode (H.pratensis), which reveals a high affinity between them. In order to determine the pathogenicity of H.avenae population in Tianzhu grasses, 2 cereal cyst nematode cysts from Kobresia and Kobresia humilis were treated at low temperature by artificial inoculation, and the eggs were inoculated to "dwarf 58" and 3000 eggs per plant. The inoculated wheat seedlings were cultured at 15 + 2 centigrade. In the room, the light was supplemental with a dysprosium lamp tube. After 14 h.1 weeks of illumination, the temperature was adjusted to 18 + 2 degrees centigrade. The soil in the PVC tube was kept certain humidity regularly, and the number of the root and rhizosphere soil cysts in the root and rhizosphere of the wheat was observed after 75 days. The results showed that the Tianzhu grass cyst nematode population could infect the wheat.
【學位授予單位】：甘肅農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S812.6

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