冬水稻田节肢动物群落调查及利用分子标记技术分析蜘蛛对弹尾虫的捕食作用

发布时间：2018-01-18 03:28

本文关键词：冬水稻田节肢动物群落调查及利用分子标记技术分析蜘蛛对弹尾虫的捕食作用　出处：《西南大学》2017年硕士论文　论文类型：学位论文

【摘要】：冬水稻田是指为保来年栽插,在水稻收割后蓄水越冬的稻田。在我国南方稻区,冬水田既是一种耕作制度,又是一种大规模的蓄水工程。在重庆等西南丘陵山区,冬水稻田作为一种特殊的农田生态系统广泛存在。本研究在对重庆市冬水稻田节肢动物三种栖息地调查的基础上,室内饲养和观察了两种优势跳虫的生活史,同时克隆了其线粒体COI基因,比较分析了两者的亲缘关系;通过室内饲养和田间取样,利用基因标记法分析了蜘蛛对弹尾虫的捕食作用。结果如下:冬水稻田系统三种生境中,总计诱捕到3,918头节肢动物,尽管个体数是田埂杂草地稻田,但3者间差异不显著(P0.05);物种类群数和Margalef物种丰富度指数均差异较小且3者间差异不显著(P0.05);Shannon-Winner多样性指数是稻田和田埂均显著大于杂草地(P0.05);Simpson多样性指数是杂草地显著大于稻田和田埂(P0.05)。猎物以腐食者天台刺齿跳虫Homidia tiantaiensis和纵纹刺齿跳虫H.socia为绝对优势种;捕食者以拟水狼蛛Pirata subpiraticus、食虫沟瘤蛛Ummeliata insecticeps、草间小黑蛛Erigonidium graminicolum、纵条蝇狮Marpissa magister和棕管巢蛛Clubiona japonicola为优势类群;这些猎物和天敌种群动态在不同栖息地结构差异明显,并随季节的变化呈现消长的变化趋势。两种优势弹尾虫的饲养结果表明,孵化期、蜕皮次数和蜕皮间隔3个指标的均值差异较小;而天台刺齿跳虫的蜕皮期要明显高于纵纹刺齿跳虫,均值分别为17.04 d和13.48 d。两种跳虫COI-658基因比对分析指出,两者间的相似度达到了91.66%,具有较近的亲缘关系。弹尾纲通用引物Col4F/Col5R仅对天台刺齿跳虫和纵纹刺齿跳虫具有扩增作用,与同域发生的其他稻虫无交叉反应,可作为冬水稻田蜘蛛捕食弹尾虫后检测该猎物的有效特异性引物;冬水稻田蜘蛛目6科11种蜘蛛均捕食弹尾虫,阳性检出率均大于40%,其中丰富度最高的几个优势种食虫沟瘤蛛、拟水狼蛛、纵条蝇狮、棕管巢蛛和草间小黑蛛的总阳性率按大小依次是51.9%、51.8%、47.4%、46.2%和45.8%。并且随月份呈递减趋势,其中当年采后季节的阳性率要显著高于翌年春季,荧光定量结果表明纵纹刺齿跳虫在拟水狼蛛、草间小黑蛛、食虫沟瘤蛛、棕管巢蛛和纵条蝇狮体内的残留拷贝数均呈现当年水稻采收后较高,随后下降并翌年春季回升的现象。实验条件下,拟水狼蛛取食纵纹刺齿跳虫后的阳性检出率随着消化时间的延长而下降,该下降曲线可用指数回归方程拟合,得到拟合方程:Y=95.6189 exp(-0.035864 X),R=0.9911,P0.0001。根据拟合曲线,该阳性检出率在0-40 h下降最快,之后减缓;最长检出时间为64h;根据拟合方程得出了该蜘蛛体内弹尾虫DNA降解50%时的消化时间DS50值为19.3 h;荧光定量PCR技术分析表明,纵纹刺齿跳虫DNA在拟水狼蛛肠道内消解主要发生在捕食后的8h内。结论:冬水稻田系统不同生境节肢动物群落结构和多样性存在明显差异;腐食者和捕食者是冬水稻田节肢动物群落的主要功能群,在采后秋季物种丰富度和多样性等要高于翌年春季;其中天台刺齿跳虫和纵纹刺齿跳虫以及蜘蛛中的拟水狼蛛、食虫沟瘤蛛、草间小黑蛛、纵条蝇狮和棕管巢蛛分别是腐食者和捕食者中的绝对优势种,对采后季节冬水田“水稻残体-腐食者-捕食者”营养级联系统起重要影响作用。这2种弹尾虫生长发育规律相似,亲缘关系较近。冬水田所有蜘蛛均能捕食弹尾虫,且捕食量较高,说明弹尾虫是该季节稻田内蜘蛛维持生存的主要猎物源;蜘蛛对弹尾虫的捕食量当年采后秋季要高于翌年春季;蜘蛛捕食弹尾虫后的消化时间主要在捕食后的8 h内,蜘蛛中肠弹尾虫DNA降解50%时的DS50值为19.3 h,该DNA残留量最长的检出时间为64h。
[Abstract]:Paddy field refers to keep planting rice harvest in the next year, after the impoundment of winter paddy field. In the south of China, winter paddy field is a farming system, and is a large-scale water storage project in Chongqing. The southwest hilly area, widely exists in paddy field as a special kind of farmland ecosystem. In this study, based on the Chongqing city winter paddy field three arthropod animal habitat survey, indoor rearing and observing the life history of two dominant Collembola, and cloning of its mitochondrial COI gene, a comparative analysis of the relationship between the two; through indoor feeding and field sampling, using genetic marker analysis of predation the spider of Collembola. The results are as follows: paddy field system in three habitats, a total of 3918 head of arthropod animal trapping, although the number of individuals is the ridge weeds of paddy fields, but there is no significant difference between the 3 groups (P0.05); species number and M Argalef species richness index showed little difference and no significant difference between the 3 (P0.05); Shannon-Winner diversity index is paddy field and ridge were significantly greater than the weed (P0.05); Simpson diversity index is significantly higher than paddy weed and ridge (P0.05). The scavengers who prey Tiantai barbed teeth tiantaiensis and longitudinal Homidia springtails tattoo teeth of Collembola H.socia was the dominant species in the predator; p.subpiraticus insecticeps Pirata subpiraticus, Ummeliata insecticeps, Erigonidium graminicolum graminicolum, marpissa Magister Marpissa Magister and Clubiona japonicola Clubiona japonicola were the dominant species; the prey population dynamics and natural enemies in the different habitat structure is obvious, change trend the changes and seasonal growth and decline. The results showed the two dominant Collembola showed that the incubation period, and the 3 molt molt times interval index mean difference Small differences; and Tiantai barbed teeth Collembola molting period was higher than that of the longitudinal tooth tattoo springtails, mean values were 17.04 D and 13.48 D. two species COI-658 gene analysis pointed out that the similarity between the two reached 91.66%, has a close genetic relationship. Collembola universal primer Col4F/ Col5R only on the roof of needling teeth Collembola and longitudinal stria barbed teeth springtails have amplification effect, without cross reaction with other sympatric occurrence of rice insect pests, as winter paddy field spiders in Collembola after the effective detection of specific primers of the prey; paddy field spiders belonging to 6 families and 11 species of spiders were prey collembolan, positive rates were greater than 40%, of which the dominant species richness insecticeps highest, p.subpiraticus, marpissa Magister, the total positive rate of Clubiona japonicola graminicolum and according to the size of the order is 51.9%, 51.8%, 47.4%, 46.2% and 45.8%. and with the month showed a decreasing trend, which then The positive rate of post harvest season was significantly higher than the next spring, quantitative results show that the longitudinal tooth tattoo springtails in p.subpiraticus, E.graminicolum, Ummeliata insecticeps, Clubiona japonicola and marpissa Magister in residual copy number showed a year after harvest of rice is higher, then decreased and the next spring rise the phenomenon. Under the experimental conditions, p.subpiraticus fed longitudinal stria barbed teeth springtails the positive rate decreased with prolonged digestion time, the decline curve can be exponential regression equation fitting, fitting equation: Y=95.6189 (Exp -0.035864 X), R=0.9911, P0.0001. according to the fitting curve, slow down after the positive rate in 0-40 h decreased most rapidly; the longest detection time was 64H; according to the fitting equation and gets the digestion time DS50 spider Collembola DNA degradation of 50% when the value is 19.3 h; fluorescence quantitative PCR analysis showed that, DNA in the longitudinal tooth tattoo to Collembola The intestinal digestion of Pirata occurs mainly in predation after 8h. Conclusion: there are obvious differences between the system of paddy field in different habitats of arthropod animal community structure and diversity; rot and predator is the main function of arthropod animal communities in winter paddy fields after harvest in autumn, species richness and diversity is higher than that of the next spring the roof; barbed teeth Collembola and longitudinal tattoo p.subpiraticus teeth and spiders in Collembola, insecticeps, E.graminicolum, marpissa Magister and Clubiona japonicola were the dominant species and predator rot in Postharvest seasons, the winter paddy rice residue "- saprophages - predator trophic cascade system play an important role. The 2 collembolan growth pattern is similar to that of a close genetic relationship. The spider can prey on all winter paddy Collembola, and predation is higher, indicating the Collembola is the season of paddy field spiders maintain The main source of prey survival; predation of spider Collembola in autumn than in spring next year after harvest; spiders in Collembola after digestion time mainly in the predator after 8 h, the spider midgut collembolan DNA degradation of 50% when the DS50 value is 19.3 h, the DNA residue in the longest check out time is 64h.

【学位授予单位】：西南大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S476;S18

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