大猿叶虫四地理种群的PCR-RFLP方法鉴别及遗传多样性分析

为了明确大猿叶虫Colaphellus bowringi Baly的遗传多样性水平及探索其种群快速鉴定的方法, 利用 7种限制性内切酶对其4个地理种群（江西龙南、江西修水、山东泰安、黑龙江哈尔滨）的线粒体COⅠ基因进行PCR-RFLP分析。结果表明: 利用AseⅠ, MboⅠ, NlaⅢ和RsaⅠ在大猿叶虫4个地理种群中均没检测到多态性; 利用AluⅠ和DraⅠ只检测到种群间的多态性; 利用HaeⅢ既检测到了种群间的多态性, 又在山东种群中检测到种群内多态性。根据酶切图谱, 共发现4种单倍型, 单倍型的特异性可以作为种群鉴别的标志。根据限制性片段共享度, 利用POPGEN 3.2计算4个种群的遗传距离, 并利用MEGA3.1进行聚类分析, 结果显示,大猿叶虫4个地理种群间的遗传距离大小与其相对地理距离的远近不相符, 其遗传变异程度与其滞育的特征也不相符。据此认为线粒体COⅠ基因的PCR-RFLP分析可以用于大猿叶虫不同地理种群的识别。     

大仓鼠种群遗传多样性的季节性变化

通过随机扩增多态性DNA（RAPD）方法,我们对河北省固安县牛驼镇王龙村附近的大仓鼠（Cricetuls triton)种群的遗传多样性进行了研究。结果显示,在209个大仓鼠个体中（春季60只,夏季45只,秋季59只,冬季45只）,共扩增出87条带,其中86．2%原条带具有多态性,4个季节种群的遗传组成存在季节变化,其多态位点数,多态位点比率,Shannon多样性指数及Nei指数的大小顺序均一致表现为：秋季种群＜冬季种群＜夏季种群＜春季种群;秋季种群的遗传多态性最低,春季最强;遗传多态性与种群密度（夹捕率,％）,之间呈极显著的负相关关系。此结果表明遗传因素与大仓鼠种群动态有密切关系,但季节间遗传多态的变化不支持Ford假说,我们认为增殖和扩散共同决定了季节间遗传多样变化的规律。     

不同地理种群小菜蛾的遗传多样性分析

采用水平切片淀粉凝胶电泳方法,分析了北京、河北、云南和武汉4个不同地理种群的小菜蛾的等位酶,得到了3个酶系统（甘油醛磷酸脱氢酶（GPD）、苹果酸酶（ME）和苹果酸脱氢酶（MDH-1,MDH-2和MDH-3）5个基因位点的资料。用Biosys2.0软件计算了不同地理种群的遗传变异指标（N、A、P、H0和He）,结果表明小菜蛾各种群内的基因多样性大于各种种群间的基因多样性（约15倍）,武汉种群小菜蛾的遗传性最大。并计算了遗传距离D和相似性系数,并由此得出聚类图,分析了小菜蛾不同地理种群间的遗传关系。     

广西红树植物桐花树种群遗传多样性分析

采用RAPD-PCR方法探讨广西3个不同生境下桐花树种群的遗传多样性和遗传分化。结果表明:3个不同生境桐花树RAPD扩增多态百分率为20.2%,3个不同生境桐花树种群两两之间的遗传距离分别为0.195、0.169、0.26,平均遗传距离为0.208。同一种群不同个体的扩增多态百分率最高为37.28%,其次为20.93%,最小的为19.32%。Shannon’s遗传多样性指数3个种群分别为0.331、0.225和0.17,其大小顺序与多态百分率的结果一致。种群内遗传多样性比率为62.3%,种群间遗传多样性比率为37.7%。说明广西3个不同生境的桐花树种群的遗传变异大部分存在种群内,种群间遗传变异较小。     

闽楠天然种群遗传多样性的RAPD分析

利用RAPD分子标记分析了闽楠主要分布区江西和福建两省的8个天然种群的遗传多样性和种群遗传分化.应用12条引物从160个植株中共检测到135个位点,其中多态位点134个.RAPD数据经Lynch-Milligan矫正后利用POPGENE软件计算出种群间遗传分化系数GST为0.373;利用Shannon多样性表型指数估算出46.4%的变异存在于种群间;分子方差分析(AMOVA)亦显示种群间变异占43.3%(P<0.001).尽管遗传变异主要存在于种群内,但闽楠种群间亦存在强烈的遗传分化,这可能与闽楠种群生境片断化、地理隔离等有关.根据闽楠的遗传变异特点,建议尽可能多地保护闽楠天然种群,对遗传多样性较高的福建西芹、浦城、明溪等种群应予以重点保护,同时收集各地闽楠遗传资源进行迁地保护,并通过种群扩繁以及合理回归自然等方式扩大种群规模、增强种群间的基因流,以维持其遗传多样性水平.     

濒危黑颈长尾雉圈养种群的RAPD遗传多样性分析

运用RAPD技术对黑颈长尾雉圈养种群的遗传多样性进行了分析。从50条随机引物中筛选出14条引物,对24个个体的基因组DNA进行了PCR扩增,从检测出的119个位点中有98个多态位点,占总位点的82.35%,标记的分子量大小范围是0.2～3kb。24个个体间的遗传距离幅度0.1597～0.4874,平均是0.2810;用软件NTsys2.10e构建了24个个体相互关系的分支图,24个个体可分为3个类群。实验表明:黑颈长尾雉圈养种群的遗传多样性水平较高,圈养种群内遗传差异性较大。     

三个地理群体赤眼鳟遗传多样性的RAPD分析

利用RAPD技术对宿鸭湖、青龙湖和丹江口水库3个野生赤眼鳟群体的遗传多样性进行分析.9个RAPD引物共获得93个扩增位点,其中多态位点56个,多态位点比例为60.22%.3个群体的多态位点比例分别为53.01%、54.12%和57.95%,遗传距离分别为0.1548、0.1613和0.1764,Shannon信息指数分别为0.2249、0.2318和0.2437.群体间遗传距离以宿鸭湖和青龙湖群体最近(0.1257),青龙湖与丹江口水库群体最远(0.1416).结果表明3个赤眼鳟群体的遗传多样性均较丰富,但群体间地理遗传分化差异并不明显.     

内蒙古典型草原羊草种群遗传分化的RAPD分析

运用 RAPD技术对内蒙古典型草原不同生境 8个羊草种群进行分析。采用 2 4个随机引物 (10 nt)在 8个种群中共检测到2 2 4个扩增片断 ,其中多态性片断 173个 ,总的多态位点百分率达 77.2 % ,特异性片断 2 2个 ,占 9.82 % ,平均每个引物扩增的DNA带数为 9.3 3条。利用 Nei指数和 Shannon指数估算了 8个种群的遗传多样性 ,并计算种群相似系数和遗传距离 ,运用UPGMA法进行聚类分析。结果表明 :羊草大部分的遗传变异存在于种群内 ,只有少部分的遗传变异存在于种群间 ,Nei指数和Shannon指数计算结果分别为 85.4%和 66.8% ;羊草不同种群的遗传多样性存在差异 ;8个羊草种群平均遗传距离为 0 .2 3 16,变异范围为 0 .1587～ 0 .2 70 0 ,说明 8个羊草种群间的遗传变异不大 ,即 :在较小地理范围内羊草的遗传分化程度较小 ;8个种群可聚为 3个类群 ,聚类结果显示生境相似的种群能够聚在一起 ,而地理距离最近的种群不一定归为一类 ,说明小范围内羊草种群间的遗传分化与地理距离不存在相关性 ,而与其生境间的相似度相关。影响遗传相似性的不是单一因子而是各种因子的综合作用 ,较小地理范围内羊草种群间的遗传分化主要是由环境的异质性所引起的     

羊草种群遗传分化的RAPD分析Ⅱ.RAPD数据的统计分析

对松嫩草原上分布的灰绿型和黄绿型羊草9个种群进行了15个引物的RAPD分析,统计结果表明,两类种群的扩增片段数和多态位点比率明显不同,黄绿型种群低于灰绿型,其值分别＜90与＞100,＜50％与＞70％,比较了7种不同统计方法据RAPD表型或基因型频率估算的种群遗传多样性,几种统计结果都揭示,黄绿型种群低于灰绿型种群,用F1s值矫正种群对Hardy-Weinberg平衡的偏离后,估算等位基因频率,通过Shannon指数和Nei指数估计羊草种群间分化分别为37．6％和35．7％,高于等位酶的分析,讨论比较了等位酶和RAPD分析结果的异同。     

中国特有种四合木种群遗传多样性的RAPD分析

采用随机扩增多态性 DNA(RAPD)技术检测了西鄂尔多斯高原特有种四合木种群 5个斑块的遗传多样性。2 2个扩增引物产生 1 1 6条带 ,Shannon信息指数和 Nei指数对 RAPD数据的分析表明 :四合木种群存在较高的遗传多样性 ,其中千里山斑块的遗传多样性和多态位点比例较高 ,石嘴山斑块的最低。遗传多样性的 86.5 %存在于斑块内 ,斑块间的遗传变异为 1 3 .5 %。遗传距离与地理距离无直接相关关系。这些结果说明 ,遗传多样性反映了四合木种群基因组 DNA存在较高的变异性 ;同时各斑块间存在一定的基因流 ,四合木各斑块可看成是处于同一种群的半隔离状态 (meta-种群 ) ,对四合木应注意保护遗传多样性丰富的 meta-种群。     

Host plant mediation of diapause induction in the cabbage beetle, Colaphellus bowringi Baly （Coleoptera： Chrysomelidae）

The cabbage beetle, Colaphellus bowringiBaly （Coleoptera： Chrysomelidae）, is a serious pest of crucifers in China, undergoing an imaginal summer and winter diapause in the soil. The effects of host plants on diapause incidence were tested in the beetle. The ratio of adults entering diapause was significantly low when they fed on the mature leaves of Chinese cabbage Shanghaiqin （Brassica chinesis var communis） than those feeding on Chinese cabbage Suzhouqin （Brassica chinesis var communis）, radish （Raphanus sativus var longipinnatus） and stem mustard （Brassicajuncea vat tumida） at 25℃ combined with 13：11 （L： D） h. Fewer adults entered diapause on young leaves compared to physiologically aged and mature radish leaves at 25℃ combined with 13：11 （L： D） h. The effect of host plant species on diapause induction was also evident under continuously dark rearing conditions or at different photoperiods. These experimental results demonstrate that host plant mediation of diapause induction exists in the cabbage beetle. However, at temperatures ≤20℃ or photoperiods of 16：8 （L： D） h combined with 25℃, all individuals entered diapause regardless of the host plants, indicating that the effects of host plants on diapause induction could be expressed only within a limited range of temperatures and photoperiods.     

Mating behavior of the cabbage beetle, Colaphellus bowringi (Coleoptera: Chrysomelidae)

Abstract  Circadian mating rhythms, mating frequency, mating duration, and the effect of mating duration on fecundity and fertility in the cabbage beetle, Colaphellus bowringi were investigated in the laboratory. Mating occurred throughout the 24-h cycle but the majority of copulations occurred in the photophase with two apparent peaks, one at 8:00 and another at 16:00. Mating frequency observations for 10 consecutive days indicated that pre-mating period of C. bowringi was about 4 days, and pairs mated an average of 5 times per day and an average of 40 times during the first 10 days. There was a negative correlation between mating frequency and mating duration during the consecutive mating. The mean duration of the first copulation (136.24 ± 4.62 min) was significantly longer than those of the second (57.87 ± 2.03 min), third (53.05 ± 2.05 min) and fourth copulation (30.86 ± 2.98 min). Fecundity showed a slight increase with increasing mating duration but no significant difference among treatments. However, fertility was significantly influenced by the mating duration in this species. Mating of 20-min duration did not produce viable eggs. The mean percentage of fertile eggs with completed mating duration (204.43 ± 18.96 min, 56.75% fertile eggs) was significantly higher than those with 60 min (39.55%) and 30 min (17.91%) mating duration, suggesting that the longer mating duration might be associated with transfer of more sperm that are used to increase the fertility of eggs.     

影响大猿叶虫交配持续时间的因素

为了探明影响大猿叶虫Colaphellus bowringi Baly交配持续时间的因素,在室内系统观察了一天中的交配起始时间、日龄、交配经历、体重、雄虫密度和温度对交配持续时间的影响.结果表明：一天中不同时间交配的成虫,其交配持续时间有显著差异,在上午8:00的交配持续时间最长,随着时间的推移,交配持续时间逐渐缩短,两者之间呈负相关.不同日龄成虫的交配持续时间有显著差异,且与成虫日龄呈明显的正相关.当雄虫与有交配经历的雌虫交配,其交配持续时间明显长于与未交配的雌虫交配,但雄虫的交配经历对交配持续时间没有影响.交配持续时间随着雌、雄体重的增加而延长,但没有显著差异.在雄虫高密度条件下的交配持续时间明显短于低密度下的交配持续时间.在22℃时成虫的交配持续时间最长,其次是25℃,28℃下最短, 温度间的交配持续时间有明显差异.这些结果表明在自然条件下大猿叶虫的交配持续时间受到许多因素的影响.     

Diapause induction and clock mechanism in the cabbage beetle, Colaphellus bowringi (Coleoptera: Chrysomelidae)

Photoperiodic control of diapause induction was investigated in the short-day species, Colaphellus bowringi, which enters summer and winter diapause as adult in the soil. Photoperiodic responses at 25 and 28 degrees C revealed a critical night length between 10 and 12 h; night lengths > or =12 h prevented diapause, whereas night lengths <12 h induced summer diapause in different degree. Experiments using non-24-h light-dark cycles showed that the duration of scotophase played an essential role in the determination of diapause. Night-interruption experiments with T=24 h showed that diapause was effectively induced by a 2-h light pulse in most scotophases; whereas day-interruption experiments by a 2-h dark break had a little effect on the incidence of diapause. The experiments of alternating short-night cycles (LD 16:8) and long-night cycles (LD 12:12) during the sensitive larval period showed that the information of short nights as well as long nights could be accumulated. Nanda-Hamner experiments showed three declining peaks of diapause at 24 h circadian intervals. Bünsow experiments showed two very weak peaks for diapause induction, one being 8 h after lights-off, and another 8 h before lights-on, but it did not show peaks of diapause at a 24 h interval. These results suggest that the circadian oscillatory system constitutes a part of the photoperiodic clock of this beetle but plays a limited role in its photoperiodic time measurement.     

Effects of thermoperiods on diapause induction in the cabbage beetle, Colaphellus bowringi (Coleoptera: Chrysomelidae)

Abstract. The effects of thermoperiods on diapause induction in continuous darkness or under a 12 : 12 h LD photoperiod were investigated in the cabbage beetle, Colaphellus bowringi Baly, a typical short‐day species. The diapause response curves both at different constant temperatures and at the thermocycle of format CT x: (24 ? x) h (16 : 28 °C) under continuously dark rearing conditions showed that the incidence of diapause depended mainly on whether or not the mean temperature was ≤20 °C or >20 °C. If the mean temperature was ≤20 °C, all individuals entered diapause; if >20 °C, the incidence of diapause declined gradually with increasing mean temperatures. The thermocycle (CT 12 : 12 h) with a series of different cryophases (8–22 °C) and thermophases (24–32 °C) under continuous darkness demonstrated a cryophase response threshold temperature of approximately 19 °C and a thermophase response threshold temperature of approximately 31 °C. Thermoperiodic amplitude (temperature difference between cryophase and thermophase) was shown to have a significant influence on diapause induction at the mean temperatures of 22, 23 and 24 °C, but not at ≥25 °C. Thermoperiodic responses under LD 12 : 12 h clearly showed that the incidence of diapause was influenced strongly by the photophase temperature. The thermoperiod under LD 12 : 12 h induced a much lower incidence of diapause than the thermoperiod with the same temperature in continuous darkness. The ecological significance of thermoperiodic induction of diapause in this species is discussed.     

大猿叶虫滞育品系和非滞育品系交配行为的比较

大猿叶虫Colaphellus bowringi Blay是一种重要的十字花科蔬菜害虫,以成虫越冬和越夏。明确大猿叶虫非滞育品系生殖力下降的现象是否与交配行为的改变相关,将促进该非滞育品系在滞育遗传、滞育调控分子机制研究中的应用。本研究在25℃和L∶D=12∶12 h条件下,测定了大猿叶虫滞育品系和非滞育品系的交配节律、交配持续时间和交配频次。结果表明,大猿叶虫滞育品系和非滞育品系交配高峰均出现在光期开始后2 h,非滞育品系的交配比率显著低于滞育品系。滞育品系和非滞育品系第1次交配持续时间无显著差异,分别为122.7±7.4 min和100.1±10.2 min;但非滞育品系的总交配持续时间显著低于滞育品系。非滞育品系的交配频次为2.2±0.3次,显著低于滞育品系的3.8±0.5次。滞育品系和非滞育品系交配行为的比较发现,大猿叶虫非滞育品系生殖力降低与其交配比率下降、交配持续时间缩短和交配频次减少显著相关,本研究为进一步利用非滞育品系为材料解析滞育遗传、滞育调控机制奠定了基础。     

Parental effects: physiological age, mating pattern, and diapause duration on diapause incidence of progeny in the cabbage beetle, Colaphellus bowringi Baly (Coleoptera: Chrysomelidae)

The cabbage beetle, Colaphellus bowringi Baly, is a short-day species that has both a facultative summer and winter adult diapause. In this study, we systematically investigated the effects of parental physiological age, mating pattern (including mating frequency and delayed mating), and diapause duration on the diapause incidence of progeny in the beetle. As parents aged, progeny diapause incidences showed considerable inter-parental variability and an abnormally varying pattern, but their predicting line was approximate to horizontal when reared under constant conditions. There was significant difference in progeny diapause incidence between multiple-mating and once-mating treatments. Higher diapause incidences in the progeny were observed in delayed-mating treatments. Progeny diapause incidences increased as diapause duration in the parental generation increased, and hibernating individuals produced more diapausing progeny than aestivating ones. Reciprocal cross tests between the post-diapause adults and non-diapause adults with no diapause history showed that the effect of parental diapause duration on the progeny diapause incidence was determined by both mothers and fathers but the mothers appeared to have a stronger effect than the fathers. Our results suggest that the parental age effect in Colaphellus bowringi might not be simply age-dependent, but may be controlled by the interaction of multiple factors, including physiological, environmental, behavioral and genetic factors. We discuss the ecological and physiological significance of these findings.     

大猿叶虫夏滞育的诱导：基于定量的光周期反应

为了探明大猿叶虫Colaphellus bowringi Baly夏滞育诱导的光周期时间测量特性, 我们通过室内实验系统比较了该虫在25℃、 不同长光照条件下,夏滞育的发生以及诱导50%个体进入夏滞育所需求的光 暗循环数。结果表明：不同长光照诱导的夏滞育比率有显著差异, 其中15 h或16 h光照诱导的滞育比率最高, 短于或长于这两个光照其滞育率均明显下降。在不同光 暗循环实验中, 14 h诱导的滞育比率均低于50%, 诱导50%个体进入夏滞育所需求的光 暗循环数在L15∶D9, L16∶D8, L17∶ D7和L18∶D6分别为2.61, 3.72, 4.64和5.92 d, 处理间存在显著差异。这些结果提示该虫夏滞育的诱导是基于定量的光周期反应。