两种色型豌豆蚜胞内共生菌研究
发布时间：2018-08-28 10:27
【摘要】：豌豆蚜(Acyrthosiphon pisum)属于半翅目蚜科,是世界范围内豆科作物及牧草的主要害虫,有红、绿两种生物性。绿色型豌豆蚜在我国分布广泛,且危害严重,而红色型目前仅在我国西部地区分布,且种群数量具有明显的增加趋势。本文研究巴克纳氏(Buchner)共生菌对两种色型豌豆蚜生长发育和种群参数的影响、两种色型豌豆蚜脱共生菌过程中体内物质含量及转氨酶活性动态变化,为更好的明确两种色型豌豆蚜种群消长动态、降低其对豆科作物及牧草的危害提供理论依据,主要研究结果如下:1.抗生素对两种色型豌豆蚜生长发育和繁殖的影响将0.2%的6种抗生素(利福平、盐酸金霉素、氯霉素、盐酸土霉素、青霉素G钾盐和硫酸链霉素)分别按同时喷洒蚕豆植株和蚜虫虫体、只喷洒蚜虫虫体和只喷洒蚕豆植株3种方式处理,获得各种处理的发育历期、平均体重、体质量差、相对日均体质量增长率和平均产蚜量等生物学参数。结果表明:3种处理方式对两种色型豌豆蚜生长发育和繁殖影响的大小顺序为:同时喷洒植株和虫体处理喷洒植株表面的处理喷洒虫体处理;6种抗生素对两种色型豌豆蚜生长发育和繁殖影响的大小顺序为:利福平盐酸土霉素盐酸金霉素硫酸链霉素氯霉素青霉素G K盐;6种抗生素经过3种不同处理方式,对两种色型豌豆蚜生长发育和繁殖影响表现为:延长发育历期,体重减轻,繁殖力下降;不同抗生素和不同处理方式对两种色型豌豆蚜生长发育和繁殖影响较大的4种组合处理顺序为:利福平同时喷洒植株和虫体处理利福平喷洒植株处理利福平喷洒虫体处理土霉素同时喷洒植株和虫体处理。2.利福平3种处理方式对两种色型豌豆蚜种群参数的影响利福平3种处理后,2种色型豌豆蚜种群净生殖率下降,平均世代周期延长,内禀增长率和周限增长率增加,种群加倍时间延长,且对种群净生殖率、平均世代周期、内禀增长率、周限增长率和种群加倍时间的影响与同色型对照差异显著。利福平3种处理对红色型和绿色型种群净生殖率、内禀增长率和种群加倍时间的影响与同色型对照差异显著。3.两种色型豌豆蚜脱共生菌利福平浓度筛选利福平植株内吸处理试验结果表明:随着利福平浓度的逐渐增大,两种色型豌豆蚜若蚜死亡率增大,世代历期缩短,繁殖力下降,成蚜体重减轻。根据蚜虫脱共生菌后的生物学参数变化,最终选择豌豆蚜脱共生菌的利福平浓度为0.2mg/ml。4.两种色型豌豆蚜脱共生菌鉴定用0.2mg/ml的6种抗生素(利福平、盐酸金霉素、氯霉素、盐酸土霉素、青霉素g钾盐和硫酸链霉素)溶液150ml进行植物内吸处理,分别在第6d和第11d取样进行豌豆蚜脱共生菌鉴定,通过分子鉴定和光学显微观察结果表明,经过6种抗生素处理后在第6d时,都不能脱掉两种色型豌豆蚜体内的buchner菌;经利福平和土霉素处理后在第11d时,两种色型豌豆蚜体内的buchner菌被脱掉。5.两种色型豌豆蚜脱共生菌过程中体内物质含量的动态变化用0.2mg/ml利福平溶液进行豌豆蚜脱共生菌处理,处理后两种色型豌豆蚜单头蚜虫蛋白含量在第4、6、8、10和12d显著低于同色型对照;处理后两种色型豌豆蚜单头可溶性糖含量第6、8、10和12d显著低于同色型对照;处理后红色型豌豆蚜单头糖原含量在第2、6、8、10和12d显著低于同色型对照,而绿色型单头蚜虫糖原含量在第6、8、10和12d显著低于对照,第2d显著高于对照;处理后两种色型豌豆蚜单头蚜虫总脂肪含量在第2d低于同色型对照,而在第4、6、8、10和12d高于同色型对照,红色型在第8d总脂肪含量与对照差异显著,而绿色型在第12d总脂肪含量与对照差异显著;处理后两种色型豌豆蚜中性脂肪含量在第2、4、6、8和10d高于同色型对照,而中性脂肪含量在第12d低于同色型对照。从物质含量角度出发,豌豆蚜脱共生菌过程是碳水化合物和蛋白质含量下降,总脂肪和中性脂肪含量升高的过程。6.两种色型豌豆蚜脱共生菌处理后能量变化用0.2mg/ml利福平溶液处理豌豆蚜后,在第8d时计算体内物质储存能量变化表明:处理后豌豆蚜体内蛋白质、可溶性糖和糖原所储存的能量显著低于同色型对照;处理后豌豆蚜体内总脂肪所储存的能量高于同色型对照,但差异不显著;处理后两种色型豌豆蚜体内总能量显著低于同色型对照。7.豌豆蚜脱共生菌过程中体内转氨酶活性动态变化研究结果表明:用0.2mg/ml利福平脱共生菌处理后,绿色型豌豆蚜谷草转氨酶(ast)活性与对照相比,第2、4、6和8d的ast酶活性高于对照,而第10和12d的处理ast酶活性低于对照;红色型ast酶活性表现为:处理后,第2、10和12d的AST酶活高于对照,其余测定时间AST酶活处理低于对照;绿色型谷丙转氨酶(ALT)酶活性只有第6d对照高于处理,其余测定时间都是对照低于处理。而红色型豌豆蚜ALT酶活性第2和4d时,对照高于处理,其余测定时间都是对照低于处理;处理后两种色型豌豆蚜谷氨酰胺合成酶(GS)酶活性都表现为:第2、4和6d都是处理高于对照,而从第8d开始,对照和处理酶活性出现交替上升变化。
[Abstract]:Acyrthosiphon Pisum is one of the main pests of leguminous crops and Forages in the world. It has red and green biological characteristics. The green pea aphid is widely distributed in China, and its harm is serious. The red pea aphid is only distributed in the western part of China, and its population has an obvious increasing trend. The effects of Buchner symbiotic bacteria on the growth and population parameters of two color-type pea aphids and the dynamic changes of substance content and aminotransferase activity in the process of de-symbiotic bacteria of two color-type pea aphids provide a theoretical basis for better understanding the population dynamics of two color-type pea aphids and reducing their damage to legume crops and forages. The main results are as follows: 1. The effects of antibiotics on the growth and reproduction of two color pea aphids were studied. Six kinds of antibiotics (rifampicin, chloramphenicol, chloramphenicol, oxytetracycline hydrochloride, penicillin G potassium salt and streptomycin sulfate) were sprayed simultaneously on broad bean plants and aphid bodies, only on aphid bodies and only on broad bean plants. The results showed that the order of the effects of the three treatments on the growth, development and reproduction of the two types of pea aphids was: spraying plants and insects simultaneously on the surface of the plants. The effects of six antibiotics on the growth, development and reproduction of two color pea aphids were in the following order: rifampicin hydrochloride oxytetracycline hydrochloride oxytetracycline hydrochloride streptomycin sulfate chloramphenicol penicillin G K salt; six antibiotics were treated in three different ways to prolong the development and reproduction of two color pea aphids. Four combinations with different antibiotics and different treatments had great effects on the growth and reproduction of two types of pea aphids, and the sequence was as follows: rifampicin sprayed on plants and rifampicin sprayed on plants at the same time; rifampicin sprayed on plants and rifampicin sprayed on insects at the same time. 2. rifampicin sprayed on plants and worms at the same time. Effects of three different treatments on the population parameters of two color-type pea aphids Rifampicin three treatments showed that the net reproductive rate of the two color-type pea aphids decreased, the average generation cycle prolonged, the intrinsic growth rate and the perimeter growth rate increased, the population doubling time prolonged, and the net reproductive rate, the average generation cycle, the intrinsic growth rate, the perimeter growth rate and the perimeter growth rate increased. The effects of rifampicin treatments on net reproductive rate, intrinsic growth rate and doubling time of red and green populations were significantly different from those of the same colour control. 3. The concentration of rifampicin screened for rifampicin in two colour types of pea aphids showed that rifampicin had significant effects on the net reproductive rate, intrinsic growth rate and doubling time of red and green populations. With the increase of rifampicin concentration, the mortality of nymphs increased, the generations shortened, the fecundity decreased, and the weight of adult aphids decreased. According to the changes of biological parameters after aphid desymbiotic bacteria, rifampicin concentration of the desymbiotic bacteria of pea aphids was 0.2 mg/ml.4. Six kinds of antibiotics (rifampicin, chloramphenicol, chloramphenicol, oxytetracycline hydrochloride, penicillin G potassium salt and streptomycin sulfate) in the solution of 150 ml were used for plant endophytic treatment. Samples were taken on the 6th and 11th days for identification of the aposymbiotic bacteria of the pea aphid. The results of molecular identification and optical microscopy showed that after six kinds of antibiotics treatment on the 6th day, The Buchner bacteria in the two types of pea aphids could not be removed. On the 11th day after treatment with rifampicin and oxytetracycline, the Buchner bacteria in the two types of pea aphids were removed. The protein content of the aphids of the two color types was significantly lower than that of the same color type on the 4th, 6th, 8th, 10th and 12th day, the soluble sugar content of the two color types was significantly lower than that of the same color type on the 6th, 8th, 10th and 12th day, and the glycogen content of the red type was significantly lower than that of the same color type on the 2nd, 6th, 8th, 10th and 12th day, while that of the green type was significantly lower than that of the same color type. The glycogen content of the aphids was significantly lower than that of the control on the 6th, 8th, 10th and 12th day, and significantly higher than that of the control on the 2nd day. The total fat content of the aphids of the two color types was lower than that of the same color type on the 2nd day, but higher than that of the same color type on the 4th, 6th, 8th, 10th and 12th day. The content of neutral fat was higher on the 2nd, 4th, 6th, 8th and 10th day than that of the control, while the content of neutral fat was lower on the 12th day. 6. Energy changes of the two kinds of decomposing bacteria after treatment with 0.2mg/ml rifampicin solution were calculated on the 8th day. The results showed that the energy stored in protein, soluble sugar and glycogen of the treated pea aphid was significantly lower than that of the control. The total energy stored in lipids was significantly lower than that of the control. 7. The dynamic changes of transaminase activity in vivo during the process of de-symbiosis of pea aphid were studied. The results showed that the green pea aphid glutamic oxaloacetic transaminase (a) was treated with 0.2 mg/ml rifampicin de-symbiosis bacteria. The activity of AST in the 2nd, 4th, 6th and 8th day was higher than that in the control, while the activity of AST in the 10th and 12th day was lower than that in the control. The ALT activity of the red pea aphid was higher than that of the control on the 2nd and 4th day, and the rest was lower than that of the control. The glutamine synthase (GS) activity of the two color pea aphids was higher than that of the control on the 2nd, 4th and 6th day, but from the 8th day, the control was higher than that of the control. And enzyme activity increased alternately.
【学位授予单位】：甘肃农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S433
本文编号：2209120
[Abstract]:Acyrthosiphon Pisum is one of the main pests of leguminous crops and Forages in the world. It has red and green biological characteristics. The green pea aphid is widely distributed in China, and its harm is serious. The red pea aphid is only distributed in the western part of China, and its population has an obvious increasing trend. The effects of Buchner symbiotic bacteria on the growth and population parameters of two color-type pea aphids and the dynamic changes of substance content and aminotransferase activity in the process of de-symbiotic bacteria of two color-type pea aphids provide a theoretical basis for better understanding the population dynamics of two color-type pea aphids and reducing their damage to legume crops and forages. The main results are as follows: 1. The effects of antibiotics on the growth and reproduction of two color pea aphids were studied. Six kinds of antibiotics (rifampicin, chloramphenicol, chloramphenicol, oxytetracycline hydrochloride, penicillin G potassium salt and streptomycin sulfate) were sprayed simultaneously on broad bean plants and aphid bodies, only on aphid bodies and only on broad bean plants. The results showed that the order of the effects of the three treatments on the growth, development and reproduction of the two types of pea aphids was: spraying plants and insects simultaneously on the surface of the plants. The effects of six antibiotics on the growth, development and reproduction of two color pea aphids were in the following order: rifampicin hydrochloride oxytetracycline hydrochloride oxytetracycline hydrochloride streptomycin sulfate chloramphenicol penicillin G K salt; six antibiotics were treated in three different ways to prolong the development and reproduction of two color pea aphids. Four combinations with different antibiotics and different treatments had great effects on the growth and reproduction of two types of pea aphids, and the sequence was as follows: rifampicin sprayed on plants and rifampicin sprayed on plants at the same time; rifampicin sprayed on plants and rifampicin sprayed on insects at the same time. 2. rifampicin sprayed on plants and worms at the same time. Effects of three different treatments on the population parameters of two color-type pea aphids Rifampicin three treatments showed that the net reproductive rate of the two color-type pea aphids decreased, the average generation cycle prolonged, the intrinsic growth rate and the perimeter growth rate increased, the population doubling time prolonged, and the net reproductive rate, the average generation cycle, the intrinsic growth rate, the perimeter growth rate and the perimeter growth rate increased. The effects of rifampicin treatments on net reproductive rate, intrinsic growth rate and doubling time of red and green populations were significantly different from those of the same colour control. 3. The concentration of rifampicin screened for rifampicin in two colour types of pea aphids showed that rifampicin had significant effects on the net reproductive rate, intrinsic growth rate and doubling time of red and green populations. With the increase of rifampicin concentration, the mortality of nymphs increased, the generations shortened, the fecundity decreased, and the weight of adult aphids decreased. According to the changes of biological parameters after aphid desymbiotic bacteria, rifampicin concentration of the desymbiotic bacteria of pea aphids was 0.2 mg/ml.4. Six kinds of antibiotics (rifampicin, chloramphenicol, chloramphenicol, oxytetracycline hydrochloride, penicillin G potassium salt and streptomycin sulfate) in the solution of 150 ml were used for plant endophytic treatment. Samples were taken on the 6th and 11th days for identification of the aposymbiotic bacteria of the pea aphid. The results of molecular identification and optical microscopy showed that after six kinds of antibiotics treatment on the 6th day, The Buchner bacteria in the two types of pea aphids could not be removed. On the 11th day after treatment with rifampicin and oxytetracycline, the Buchner bacteria in the two types of pea aphids were removed. The protein content of the aphids of the two color types was significantly lower than that of the same color type on the 4th, 6th, 8th, 10th and 12th day, the soluble sugar content of the two color types was significantly lower than that of the same color type on the 6th, 8th, 10th and 12th day, and the glycogen content of the red type was significantly lower than that of the same color type on the 2nd, 6th, 8th, 10th and 12th day, while that of the green type was significantly lower than that of the same color type. The glycogen content of the aphids was significantly lower than that of the control on the 6th, 8th, 10th and 12th day, and significantly higher than that of the control on the 2nd day. The total fat content of the aphids of the two color types was lower than that of the same color type on the 2nd day, but higher than that of the same color type on the 4th, 6th, 8th, 10th and 12th day. The content of neutral fat was higher on the 2nd, 4th, 6th, 8th and 10th day than that of the control, while the content of neutral fat was lower on the 12th day. 6. Energy changes of the two kinds of decomposing bacteria after treatment with 0.2mg/ml rifampicin solution were calculated on the 8th day. The results showed that the energy stored in protein, soluble sugar and glycogen of the treated pea aphid was significantly lower than that of the control. The total energy stored in lipids was significantly lower than that of the control. 7. The dynamic changes of transaminase activity in vivo during the process of de-symbiosis of pea aphid were studied. The results showed that the green pea aphid glutamic oxaloacetic transaminase (a) was treated with 0.2 mg/ml rifampicin de-symbiosis bacteria. The activity of AST in the 2nd, 4th, 6th and 8th day was higher than that in the control, while the activity of AST in the 10th and 12th day was lower than that in the control. The ALT activity of the red pea aphid was higher than that of the control on the 2nd and 4th day, and the rest was lower than that of the control. The glutamine synthase (GS) activity of the two color pea aphids was higher than that of the control on the 2nd, 4th and 6th day, but from the 8th day, the control was higher than that of the control. And enzyme activity increased alternately.
【学位授予单位】：甘肃农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S433
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