棉蚜在花椒上的种群动态及花椒香豆素对棉蚜生化特性的影响
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摘要
重庆是我国主要的青花椒生产基地,种植面积已达76万亩,居全国前列。但随着花椒生产的不断发展,病虫害发生危害日趋严重,其中以棉蚜的危害最为猖獗。本论文对花椒园内棉蚜周年发生的种群动态、不同温度下棉蚜在花椒上的生长发育以及花椒次生物质对棉蚜的抗生作用等进行了研究,并在此基础上探讨了花椒香豆素对棉蚜主要糖转移酶和解毒酶的影响。主要研究结果如下:
1.花椒园棉蚜自然种群动态研究
周年调查表明,棉蚜在花椒上的数量动态基本上呈“双峰形”,第一个高峰发生在5月初至6月底,另一个发生在9月份,以第一个高峰虫口数量为主,且若蚜和成蚜出现的高峰期基本一致,均在南向方位分布量最大。聚集度指标分析表明,棉蚜在越冬期平均密度较低,种群为均匀分布,在其它时段则均为聚集分布,并得出棉蚜个体群大小为4.5头;而扩散和聚集趋势分析表明,棉蚜在花椒上表现出扩散→聚集→再扩散→再聚集的趋势。并确定了不同虫口密度下的最适理论抽样数及一定防治指标下的序贯抽样模型:春蚜期T_((n))=5n±4.5709n~(1/2),伏蚜期T_((n))=50n±24.8412n~(1/2),n为序贯抽样数。
2.温度对棉蚜生长发育的影响
在14℃、18℃、21℃、24℃、27℃和30℃六个恒温条件下,以九叶青花椒为寄主,研究了温度对棉蚜的存活率、平均寿命及繁殖力的变化。结果表明:14℃时,若蚜存活率和成蚜产仔量最小,分别只有66.94%和13.5687头,净生殖率R_0、内禀增长率γ_m和周限增长率λ均最小。18℃时,棉蚜的成虫平均寿命最长,死亡高峰日出现最晚,随着温度上升,死亡高峰日提前,30℃时仅为5.28天。18℃～21℃时,棉蚜在花椒上的繁殖力最大,产仔量分别为35.8216头和35.0802头,随着温度上升产仔数反而下降,30℃仅为16.87头,且30℃时日产仔量曲线下降最快。21℃～24℃时棉蚜存活率最高,均达到90%以上。27～30℃高温使其发育历期缩短,而内禀增长率γ_m却显著增加,30℃时γ_m值达到0.3364,表明高温能导致短期内棉蚜种群剧增。运用净生殖率R_0来评价棉蚜在花椒上的生存和繁殖能力,得出21.7℃为最适温度。
3.花椒几种次生物质对棉蚜的抗生作用
采用人工营养液饲养蚜虫技术,分析花椒精油单体成分a-蒎烯、β-蒎烯和花椒香豆素对棉蚜的抗生作用。结果表明,三种次生物质均对棉蚜有一定的抗生性。其中β-蒎烯对棉蚜的抗性作用最小,仅表现很弱的忌避性;a-蒎烯则主要在低浓度(0.015%～0.045%)时表现较强的忌避性和拒食性,当浓度升高到0.060%时抗性作用反而减小:而香豆素在低浓度时对棉蚜的抗性不明显,但在高浓度(＞0.060%)对棉蚜的抗性显著增加,其中以致毒作用为主,对棉蚜的蜜露分泌也有显著影响。
4.花椒香豆素对棉蚜糖转移酶系的影响
通过用酶标仪测定了花椒香豆素对棉蚜主要糖转移酶活性的影响。结果表明,尽管不同浓度香豆素处理下的棉蚜淀粉酶、蔗糖酶和海藻糖酶活力和比活力随处理浓度的不同而变化,但与对照相比均无显著差异。可见,香豆素对棉蚜糖转移酶没有明显作用,说明香豆素不影响棉蚜对糖的转化和能量利用。
5.花椒香豆素对棉蚜解毒酶系的影响
分析了不同浓度的香豆素对棉蚜羧酸酯酶、酸性磷酸酯酶、碱性磷酸酯酶以及谷胱甘肽S-转移酶的离体抑制作用,结果表明,香豆素对这四种解毒酶均有显著的抑制作用。其中,0.06%香豆素能显著抑制棉蚜CarE的活性,抑制率达到52.55%,但随着处理浓度升高,香豆素对CarE的抑制作用反而减小;对ACP和ALP表现出的抑制作用则相对较小,而且ACP和ALP的活性均随香豆素处理浓度的升高而降低;香豆素对棉蚜GSTs的抑制作用则最强,浓度为0.02%处理时抑制率就达到了31.1%,当浓度升高到0.1%时,抑制率达到最大78.76%。可见,香豆素对棉蚜的不良影响主要是使棉蚜解毒能力下降。
The main production base of Green Prickley ash in China is Chongqing,of which planting area has already amount to 760,000 mu.However,with the continuous development of Prickley ash production,the harms of pests and diseases are prone to be more and more serious,especially,Aphis gossypii.In this study,we investigated the annual population dynamics of Aphis gossypii in Prickly ash orchard,the growth-and-development of Aphis gossypii on Prickley ash under different temperatures and the resistance of secondary metabolites from Prickly ash to Aphis gossypii. Forthmore,we analyzed the effect of Prickly ash coumarin on the sugar invert enzymes and detoxification enzymes of Aphis gossypii.The main results were as follows:
1.The natural population dynamics of Aphis gossypii
Systematic investigations were carried out on the dynamics of natural population of Aphis gossypii in Prickly ash orchard.The result indicated that the population quantity of the insect was obviously increased from the early May to the late June,then decreased till September,and increased again,the densities of adult and larva at the position in South direction were markedly higher than other directions.Spatial distribution information showed that the population was uniform pattern during the winter,but aggregate pattern at other times,the individual group was 4.5 per leaf,and the aphid population traded off during the growth period of Prickly ash.The most suitable theoretical sampling quantity at different densities and the sequential sampling model were presented:T_(n)= 5n±4.5709n~(1/2)(Spring Aphids),T_(n)= 50n±24.8412n~(1/2)(Summer Aphids),n was the sequential sampling quantity.
2.The influence of temperature on the growth-and-development of Aphis gossypii
Analyzing the survival rate,mean longevity and fecundity of Aphis gossypii on Prickley ash at 6 constant temperatures 14℃,18℃,21℃,24℃,27℃and 30℃,we found:At 14℃,the survival rate of larva and the reproduction rate of adult were 66.94%and 13.5687 respecively,the reproductive rate(R_0).the intrinsic rate of increase(r_m)and the finite rate of increase(λ)were the lowest.At 18℃,the mean longevity of adult was the longest,and the date of maxiun mortality was the latest among these six temperatures.As the temperature increasing,the date of maxiun mortality was advanced,the shortest date of maxiun mortality was 5.28d at 30℃.At 18℃～21℃,the fecundities of cotton aphid were the highest on Prickley ash,and the reproduction quantities per female were 35.8216 and 35.0802 respecively.The survival rate of larva(92.33%and 90.14%)at 21℃and 24℃were significantly greater than those at other temperatures.The developmental duration was shortened at high temperature(27～30℃),but r_m increased significantly at 30℃ (0.3364),which implied that high temperature can cause the population increasing observably in short times.According to R_0,21.7℃was considered to be the most suitable temperature for Aphis gossypii.
3.The anti-cotton aphid fiction of secondary metabolites from prickly ash
By using the artificial rearing technique of aphids,we evaluated the resistance of a-pinene,β-pinene and coumarin from prickly ash to Aphis gossypii.The result suggested that they all had anti-cotton aphid action.Among the three kinds of secondary metabolites,theβ-pinene's resistance was the weakest.The a-pinene had strong repellent and antifeed action when its concentration was low(0.015%to 0.045%),while the concentration increased to 0.060%,the anti-cotton aphid action of a-pinene decreased on the contrary.Resistance of coumarin to Aphis gossypii was not obvious in low concentration,but in high concentration(＞0.060%)the resistance increased significantly, mainly represented toxic effects.
4.The influence of prickly ash coumarin to the sugar invert enzymes of Aphis gossypii
To further understand resistant mechanism of prickly ash coumarin to aphid,the activities of sugar invert enzymes of Aphis gossypii were detected with microtiter plate reader after adding different dosage of coumarin.The result showed there was no marked difference in the activities of amylase,sucrase and trehalase.It indicated that coumarin had not obvious effect on the transition of sugar and energy of Aphis gossypii.
5.The influence of prickly ash coumarin to the detoxification enzymes of Aphis gossypii
The impact of coumarin on the activity of carboxylesterase(CarE),acid phosphatase(ACP), alkaline phosphatase(ALP)and glutathione S-transferase(GST)in Aphis gossypii was measured. The result revealed that coumarin strongly inhibited the activity of CarE when its concentration was 0.06%,the inhibition rate was 52.55%,but the inhibition decreased when its concentration incresed. The inhibition of coumarin to ACP and ALP was not great,and the activities all decreased as the concentration of coumarin increased.The inhibition of coumarin to GST was the strongest of all,the inhibition rate was 31.1%when its concentration was 0.02%,and the inhibition rate was the highest (78.76%)when its concentration was 0.1%.The results implied that coumarin could reduce the antidotal ability of Aphis gossypii by inhibiting the activity of detoxification enzymes.
1.花椒园棉蚜自然种群动态研究
周年调查表明,棉蚜在花椒上的数量动态基本上呈“双峰形”,第一个高峰发生在5月初至6月底,另一个发生在9月份,以第一个高峰虫口数量为主,且若蚜和成蚜出现的高峰期基本一致,均在南向方位分布量最大。聚集度指标分析表明,棉蚜在越冬期平均密度较低,种群为均匀分布,在其它时段则均为聚集分布,并得出棉蚜个体群大小为4.5头;而扩散和聚集趋势分析表明,棉蚜在花椒上表现出扩散→聚集→再扩散→再聚集的趋势。并确定了不同虫口密度下的最适理论抽样数及一定防治指标下的序贯抽样模型:春蚜期T_((n))=5n±4.5709n~(1/2),伏蚜期T_((n))=50n±24.8412n~(1/2),n为序贯抽样数。
2.温度对棉蚜生长发育的影响
在14℃、18℃、21℃、24℃、27℃和30℃六个恒温条件下,以九叶青花椒为寄主,研究了温度对棉蚜的存活率、平均寿命及繁殖力的变化。结果表明:14℃时,若蚜存活率和成蚜产仔量最小,分别只有66.94%和13.5687头,净生殖率R_0、内禀增长率γ_m和周限增长率λ均最小。18℃时,棉蚜的成虫平均寿命最长,死亡高峰日出现最晚,随着温度上升,死亡高峰日提前,30℃时仅为5.28天。18℃～21℃时,棉蚜在花椒上的繁殖力最大,产仔量分别为35.8216头和35.0802头,随着温度上升产仔数反而下降,30℃仅为16.87头,且30℃时日产仔量曲线下降最快。21℃～24℃时棉蚜存活率最高,均达到90%以上。27～30℃高温使其发育历期缩短,而内禀增长率γ_m却显著增加,30℃时γ_m值达到0.3364,表明高温能导致短期内棉蚜种群剧增。运用净生殖率R_0来评价棉蚜在花椒上的生存和繁殖能力,得出21.7℃为最适温度。
3.花椒几种次生物质对棉蚜的抗生作用
采用人工营养液饲养蚜虫技术,分析花椒精油单体成分a-蒎烯、β-蒎烯和花椒香豆素对棉蚜的抗生作用。结果表明,三种次生物质均对棉蚜有一定的抗生性。其中β-蒎烯对棉蚜的抗性作用最小,仅表现很弱的忌避性;a-蒎烯则主要在低浓度(0.015%～0.045%)时表现较强的忌避性和拒食性,当浓度升高到0.060%时抗性作用反而减小:而香豆素在低浓度时对棉蚜的抗性不明显,但在高浓度(＞0.060%)对棉蚜的抗性显著增加,其中以致毒作用为主,对棉蚜的蜜露分泌也有显著影响。
4.花椒香豆素对棉蚜糖转移酶系的影响
通过用酶标仪测定了花椒香豆素对棉蚜主要糖转移酶活性的影响。结果表明,尽管不同浓度香豆素处理下的棉蚜淀粉酶、蔗糖酶和海藻糖酶活力和比活力随处理浓度的不同而变化,但与对照相比均无显著差异。可见,香豆素对棉蚜糖转移酶没有明显作用,说明香豆素不影响棉蚜对糖的转化和能量利用。
5.花椒香豆素对棉蚜解毒酶系的影响
分析了不同浓度的香豆素对棉蚜羧酸酯酶、酸性磷酸酯酶、碱性磷酸酯酶以及谷胱甘肽S-转移酶的离体抑制作用,结果表明,香豆素对这四种解毒酶均有显著的抑制作用。其中,0.06%香豆素能显著抑制棉蚜CarE的活性,抑制率达到52.55%,但随着处理浓度升高,香豆素对CarE的抑制作用反而减小;对ACP和ALP表现出的抑制作用则相对较小,而且ACP和ALP的活性均随香豆素处理浓度的升高而降低;香豆素对棉蚜GSTs的抑制作用则最强,浓度为0.02%处理时抑制率就达到了31.1%,当浓度升高到0.1%时,抑制率达到最大78.76%。可见,香豆素对棉蚜的不良影响主要是使棉蚜解毒能力下降。
The main production base of Green Prickley ash in China is Chongqing,of which planting area has already amount to 760,000 mu.However,with the continuous development of Prickley ash production,the harms of pests and diseases are prone to be more and more serious,especially,Aphis gossypii.In this study,we investigated the annual population dynamics of Aphis gossypii in Prickly ash orchard,the growth-and-development of Aphis gossypii on Prickley ash under different temperatures and the resistance of secondary metabolites from Prickly ash to Aphis gossypii. Forthmore,we analyzed the effect of Prickly ash coumarin on the sugar invert enzymes and detoxification enzymes of Aphis gossypii.The main results were as follows:
1.The natural population dynamics of Aphis gossypii
Systematic investigations were carried out on the dynamics of natural population of Aphis gossypii in Prickly ash orchard.The result indicated that the population quantity of the insect was obviously increased from the early May to the late June,then decreased till September,and increased again,the densities of adult and larva at the position in South direction were markedly higher than other directions.Spatial distribution information showed that the population was uniform pattern during the winter,but aggregate pattern at other times,the individual group was 4.5 per leaf,and the aphid population traded off during the growth period of Prickly ash.The most suitable theoretical sampling quantity at different densities and the sequential sampling model were presented:T_(n)= 5n±4.5709n~(1/2)(Spring Aphids),T_(n)= 50n±24.8412n~(1/2)(Summer Aphids),n was the sequential sampling quantity.
2.The influence of temperature on the growth-and-development of Aphis gossypii
Analyzing the survival rate,mean longevity and fecundity of Aphis gossypii on Prickley ash at 6 constant temperatures 14℃,18℃,21℃,24℃,27℃and 30℃,we found:At 14℃,the survival rate of larva and the reproduction rate of adult were 66.94%and 13.5687 respecively,the reproductive rate(R_0).the intrinsic rate of increase(r_m)and the finite rate of increase(λ)were the lowest.At 18℃,the mean longevity of adult was the longest,and the date of maxiun mortality was the latest among these six temperatures.As the temperature increasing,the date of maxiun mortality was advanced,the shortest date of maxiun mortality was 5.28d at 30℃.At 18℃～21℃,the fecundities of cotton aphid were the highest on Prickley ash,and the reproduction quantities per female were 35.8216 and 35.0802 respecively.The survival rate of larva(92.33%and 90.14%)at 21℃and 24℃were significantly greater than those at other temperatures.The developmental duration was shortened at high temperature(27～30℃),but r_m increased significantly at 30℃ (0.3364),which implied that high temperature can cause the population increasing observably in short times.According to R_0,21.7℃was considered to be the most suitable temperature for Aphis gossypii.
3.The anti-cotton aphid fiction of secondary metabolites from prickly ash
By using the artificial rearing technique of aphids,we evaluated the resistance of a-pinene,β-pinene and coumarin from prickly ash to Aphis gossypii.The result suggested that they all had anti-cotton aphid action.Among the three kinds of secondary metabolites,theβ-pinene's resistance was the weakest.The a-pinene had strong repellent and antifeed action when its concentration was low(0.015%to 0.045%),while the concentration increased to 0.060%,the anti-cotton aphid action of a-pinene decreased on the contrary.Resistance of coumarin to Aphis gossypii was not obvious in low concentration,but in high concentration(＞0.060%)the resistance increased significantly, mainly represented toxic effects.
4.The influence of prickly ash coumarin to the sugar invert enzymes of Aphis gossypii
To further understand resistant mechanism of prickly ash coumarin to aphid,the activities of sugar invert enzymes of Aphis gossypii were detected with microtiter plate reader after adding different dosage of coumarin.The result showed there was no marked difference in the activities of amylase,sucrase and trehalase.It indicated that coumarin had not obvious effect on the transition of sugar and energy of Aphis gossypii.
5.The influence of prickly ash coumarin to the detoxification enzymes of Aphis gossypii
The impact of coumarin on the activity of carboxylesterase(CarE),acid phosphatase(ACP), alkaline phosphatase(ALP)and glutathione S-transferase(GST)in Aphis gossypii was measured. The result revealed that coumarin strongly inhibited the activity of CarE when its concentration was 0.06%,the inhibition rate was 52.55%,but the inhibition decreased when its concentration incresed. The inhibition of coumarin to ACP and ALP was not great,and the activities all decreased as the concentration of coumarin increased.The inhibition of coumarin to GST was the strongest of all,the inhibition rate was 31.1%when its concentration was 0.02%,and the inhibition rate was the highest (78.76%)when its concentration was 0.1%.The results implied that coumarin could reduce the antidotal ability of Aphis gossypii by inhibiting the activity of detoxification enzymes.
引文
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