Comparative Analysis of Genetic Diversity Among Indian Populations of Scirpophaga incertulas by ISSR-PCR and RAPD-PCR

Genetic variation between 28 Indian populations of the rice pest, Scirpophaga incertulas was evaluated using inter-simple sequence repeats (ISSR)-PCR assay. Nine SSR primers gave rise to 79 amplification products of which 67 were polymorphic. A dendrogram constructed from this data indicates that there is no geographical bias to the clustering and that gene flow between populations appears to be relatively unrestricted, substantiating our earlier conclusion based on the RAPD (random amplified polymorphic DNA) data. The dendrograms obtained using each of these marker systems were poorly correlated with each other as determined by Mantel's test for matrix correlation. Estimates of expected heterozygosity and marker index for each of these marker systems suggests that both these marker systems are equally efficient in determining polymorphisms. Matrix correlation analyses suggest that reliable estimates of genetic variation among the S. incertulas pest populations can be obtained by using RAPDs alone or in combination with ISSRs, but ISSRs alone cannot be used for this purpose.     

闽北地区四代三化螟发生动态Fuzzy预测模型研究

根据Fuzzy数学原理和三化螟生物学特性,组建了四代三化螟发生动态综合预测模型经对福建省将乐县16年四代三化螟发生期、发生量回报及1995年的预报,拟合率及正确率均达100％。     

Population decrease of Scirpophaga incertulas Walker (Lepidoptera Pyralidae) under climate warming

Scirpophaga incertulas Walker is an important agricultural pest in Asia. Only few studies are available on its long-term population dynamics under climate warming. In this study, we used the linear and generalized additive models (GAMs) to analyze the historical dataset of >50 years on this pest at Xinfeng County of Jiangxi Province, China. The main objective of this study was to explore the effects of density (delayed) dependence and minimum annual temperature (MAT), which indirectly reflects climate warming, on the population dynamics of this pest. We found that both density dependence and MAT have significant influence on the annual population growth rate. The GAMs had relatively better applicability to the dataset than the linear models. Nonparametric model provided satisfactory goodness-of-fit (R(2) > 0.5). At Xinfeng County, the MAT had a significant effect on the annual population growth rate of S. incertulas. The annual population growth rate of S. incertulas decreased with increase in MAT. Therefore, S. incertulas population becomes smaller and smaller in Southern China due to climate warming. The current study has two contributions: (1) providing a suitable method for predicting the annual population growth rate of S. incertulas, and (2) demonstrating that climate warming could decrease the S. incertulas population.     

人工神经网络在三化螟预测中的应用

本应用神经网络对闽北四代三化螟的发生情况进行了预测,三化螟的预报可视作高维空间的非线性分类问题。利用BP网络的反向传播算法．我们对三化螟种群发生趋势进行预测,并获得了满意的结果。     

转 Bt 基因水稻对三化螟自然种群的影响

为了了解水稻重要害虫三化螟 Scirpophaga incertulas（Walker）对转 Bt 基因水稻产生抗性的潜在风险,本文利用自然种群生命表,研究了第 3 代三化螟在转Bt基因杂交稻汕优 63 及非转 Bt 基因杂交稻汕优 63 上的种群参数。结果表明：在 Bt 稻上1～2 龄幼虫的存活率为 4.03%,极显著低于非转基因 SY63（55.76%）;Bt 稻上幼虫为害率为1.75%,极显著低 于非转基因 SY63（5.98%）;Bt 稻上存活的幼虫发育历期明显长于非转 Bt SY63。     

数学分析方法在水稻螟虫测报上的应用研究

本利用福建省将乐县1963-1994年测报积累的观察赍料和气象资料,建立了越冬代二化螟、三化螟蛾高峰逐步回归顶测模型．Fuzzy分析预报模型,四代三化螟发生期发生量璜测模型。     

Varietal resistance, population dynamics and timing insecticidal application with peak oviposition by Scirpophaga incertulas (Walker) (Lepidoptera: Pyralidae) on rice

The flight activity of yellow stem borer Scirpophaga incertulas (Walker) (Lepidoptera: Pyralidae) peaked in the months of April-May, May-June, August-September and October. The number of egg masses and the number of adults attracted to light sources were the indicators of S. incertulas flight activity. The rice varieties TKM6, IR22, IR60, IR66 and IR74 were infested at 7, 10, 12 and 16 wk after planting with 5, 10, 20 and 40 neonates of S. incertulas. All varieties except IR66 were susceptible to dead heart damage by S. incertulas. When the rice varieties TKM6, BPIRi2, BPIRi4, IR22, IR36, IR60, IR66 and IR74 were treated with carbofuran insecticide at the time of peak oviposition by S. incertulas in the field, the dead heart damage on all the varieties was significantly reduced in comparison with the untreated plots. Indiscriminate routine insecticidal treatments (fixed schedule) can be replaced by a treat-when-necessary schedule based on the population dynamics of S. incertulas.     

Frequency of alleles conferring resistance to a Bacillus thuringiensis toxin in a Philippine population of Scirpophaga incertulas (Lepidoptera: Pyralidae)

Using the F2 screen methodology, we estimated the frequency of alleles conferring resistance to the Cry1Ab toxin of Bacillus thuringiensis Berliner in a Philippine population of the stem borer Scirpophaga incertulas (Walker). Evaluation of >450 isofemale lines for survival of F2 larvae on cry1Ab plants did not detect the presence of an allele conferring a high level of resistance. The frequency of such an allele in the sampled population was conservatively estimated to be <3.6 x 10(-3) with 95% confidence and a detection probability of 94%. However, there was evidence of the presence of alleles conferring partial resistance to Cry1Ab. The frequency of alleles for partial resistance was estimated as 4.8 x 10(-3) with a 95% CI between 1.3 x 10(-3) and 1.04 x 10(-2) and a detection probability of 94%. Our results suggest that the frequency of alleles conferring resistance to Cry1Ab in the population of S. incertulas sampled is not too high to preclude successful implementation of the high dose/refuge resistance management strategy.     

Physiological time model of Scirpophaga incertulas (Lepidoptera: Pyralidae) in rice in Guandong Province, People's Republic of China

Scirpophaga incertulas (Walker) (Lepidoptera: Pyralidae) is autochthonous and monophagous on rice, Oryza spp., which favors the development of a physiological time model using degree-days (degrees C) to establish a well defined window during which adults will be present in fields. Model development of S. incertulas adult flight phenology used climatic data and historical field observations of S. incertulas from 1962 through 1988. Analysis of variance was used to evaluate 5,203 prospective models with starting dates ranging from 1 January (day 1) to 30 April (day 121) and base temperatures ranging from -3 through 18.5 degrees C. From six candidate models, which shared the lowest standard deviation of prediction error, a model with a base temperature of 10 degrees C starting on 19 January was selected for validation. Validation with linear regression evaluated the differences between predicted and observed events and showed the model consistently predicted phenological events of 10 to 90% cumulative flight activity within a 3.5-d prediction interval regarded as acceptable for pest management decision making. The degree-day phenology model developed here is expected to find field application in Guandong Province. Expansion to other areas of rice production will require field validation. We expect the degree-day characterization of the activity period will remain essentially intact, but the start day may vary based on climate and geographic location. The development and validation of the phenology model of the S. incertulas by using procedures originally developed for pecan nut casebearer, Acrobasis nuxvorella Neunzig, shows the fungibility of this approach to developing prediction models for other insects.     

水稻种植密度和生育期对三化螟卵天敌种群及其效益的影响

研究结果表明,水稻种植密度和生育期对三化螟Scirpophaha incertulas卵的天敌种群及其捕食串和寄生率有显著的影响。捕食性天敌和某些寄生蜂的种群密度随种植密度的增加和生育期的增长而增加,但啮小蜂和赤眼蜂对种植密度的反应不明显。螟卵捕食串随种植密度的增加和生育期的增长而增加。但螟卵寄生率则相反：种植密度低、生育期早,卵寄生率高,且差异显著。种植密度和生育期不仅影响了单种寄生蜂的寄生率,而且也影响了螟卵寄生蜂种类的多少。在种植密度低的地块和水稻生长初期,卵块通常有3种寄生蜂出现,而在密度高的地块和生长末期,卵块通常没有寄生蜂或仅有赤眼蜂出现。虽然卵捕食和卵寄生都是三化螟卵的重要控制因素,但由天敌所造成的卵死亡率则随种植密度的增加或生育期的延长而下降。     

Random amplified polymorphic DNA markers for discriminating Cochliomyia hominivorax from C. macellaria (Diptera: Calliphoridae)

The screwworm, Cochliomyia hominivorax (Coquerel), is one of the most important pests of livestock in the Western Hemisphere. During early immature stages it is morphologically very similar (first instars are virtually indistinguishable) to the secondary screwworm, C. macellaria (Fabricius). Here, the utility of the random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) was explored as a technique for developing molecular genetic markers for these two species. Of the 120 arbitrary primers screened, 21 primers produced markers that were further investigated. Seven of the 21 primers produced clear and reproducible markers that were tested with DNA of five individuals from four populations of each species; five of these primers showed 12 RAPD markers that differentiated the species in all populations. Analyses of data from these seven primers also suggested that intraspecific polymorphisms exist that could be useful in distinguishing populations of screwworms. Some population genetic tools, such as genetic distance, cluster analysis and bootstrapping, were used to statistically explore these polymorphisms. The resulting statistics showed 100% support for the ability of RAPD-PCR to discriminate between the two species. Bootstrapping with data from one of the genetic distance calculations produced a tree with all individual screwworms in the correct populations, indicating that RAPD-PCR has promise for displaying intraspecific genetic variation that could be used in establishing the general geographic origin of screwworm samples.     

三化螟幼虫的田间分布型

通过采用聚集度指标的方法,对三化螟幼虫在稻田的空间格局进行了系统的研究。测定了幼虫个体群大小、分析聚集原因,为田间抽样、《查定》、及防治提供了依据。     

Analysis of Genetic Diversity of Chukar Partridge (Alectoris chukar) Populations in Khorasan-e-Razavi Province of Iran by RAPD-PCR

RAPD markers were used to investigate population genetic parameters of an endangered partridge, Alectoris chukar, in four areas of Iran, as a part of a genetic conservation program. The aim of this study was to analyze the genetic similarity among these populations. Blood samples from 75 birds were used for DNA extraction and RAPD-PCR analysis of 67 loci, with 28 polymorphic bands (41.79%). The populations of Kalat-e-Nader and Mashhad were found to be closely related, as were the Torbat-e-Jaam and the Quchan populations. Mean heterozygosity for all populations was 0.4405 ± 0.0755. The results indicate that chukar partridge genetic diversity in Khorasan-e-Razavi province is sufficient and the amount of gene flow among populations is acceptable.     

再生稻与三化螟越冬幼虫的关系的研究

在华南,秋季水稻收获后,留在田间的一些稻桩,既含有再生稻株,亦藏蛰伏的三化螟幼虫,当再生稻桩率为14％～19％时,其中藏有的三化螟幼虫占该虫总数的47％～65％。由于三化螟得到再生稻株的保护,在整个越冬期间藏于再生株内的蝗虫死亡率偏低,至多为非再生株同时的50％,再生株百株活虫数为26～42头,而非再生株内仅为1～7头。     

Variation in performance on cry1Ab-transformed and nontransgenic rice varieties among populations of Scirpophaga incertulas (Lepidoptera: Pyralidae) from Luzon Island, Philippines

We quantified variation in performance under greenhouse conditions among seven populations of Scirpophaga incertulas (Walker) from Luzon Island, Philippines, on three rice varieties: 'IR58' transformed with the cry1Ab gene from Bacillus thuringiensis Berliner, and nontransgenic IR58 and IR62. On IR62, S. incertutas performance did not differ among provinces for any of the 10 parameters measured, but there was a significant effect of town within province for one parameter, 20-d-old larval weight. Larval survival after 48 h on cy1Ab-transformed IR58 did not differ significantly among provinces, but did differ significantly among towns within a province. There was no geographic variation in larval survival after 48 h on control plants of IR58. Surviving insects from the cry1Ab-transformed IR58 were transferred to IR62 to complete development. There was no geographic variation in the percentage of insects completing development to adult emergence and the time required by the transferred female insects to complete development. However, there was variation among provinces in male developmental time. The absence of geographic variation on nontransgenic IR58 and the very limited variation on IR62 indicated that there was little variation in general vigor among the S. incertulas populations and thus that the variation in performance oil cry1Ab-transformed IR58 was probably attributable to differences in susceptibility to Cry1Ab.     

Development and characterization of microsatellite markers for rice leaffolder, Cnaphalocrocis medinalis (Guenée) and cross-species amplification in other Pyralididae

Rice leaffolder, Cnaphalocrocis medinalis (Guenée), is a destructive and widespread pest on rice. In this study, 20 microsatellite markers were isolated and characterized from C. medinalis partial genomic libraries using the method of fast isolation by AFLP of sequence containing repeats. Of these markers, 18 markers displayed polymorphisms. Polymorphisms were evaluated in 48 individuals from two natural populations. The number of alleles per locus ranged from 2 to 15, and the expected and observed heterozygosities ranged from 0.324 to 0.934 and from 0.304 to 0.917, respectively. Cross-species amplification was also performed to test the transferability of the 20 microsatellite markers and a moderate level of cross amplication was observed across the three species of Pyralididae (26.67 %). These microsatellite loci would facilitate the future study on population genetics and molecular genetics of rice leaffolder and would also be useful for study in Chilo suppressalis, Scirpophaga incertulas and Pyrausta nubilalis.     

Genetic variation in an introduced aphid pest (Metopolophium dirhodum) in New Zealand and relation to individuals from europe

RAPD-PCR was used to determine the genetic variation of Metopolophium dirhodum collected in a winter wheat field and in a nearby 2.5-m-high suction trap at Lincoln, New Zealand. Over three collection dates, five distinct genotypes were identified, using two primers (OPK16 and OPC09) independently. There was a significant temporal effect on the ratio of genotypes in populations collected in the field. There was no significant spatial aggregation or association of these genotypes in the field. Two of the genotypes present in the field were also detected in the suction trap sample. Using a higher resolution method of RAPD-PCR (with the Stoffel fragment of Taq polymerase), a total of 124 genotypes were distinguished from 142 individuals collected from Scotland and New Zealand. The Jaccard similarity index ( S ) was used to measure similarity between individual aphids within and between populations from both hemispheres. All populations were very diverse ( S < 0.33). However, at similar crop growth stages, M. dirhodum was significantly more diverse in Scotland than in New Zealand. The results are discussed in relation to the value of monitoring aphid flights for pest forecasting, and in terms of the most appropriate RAPD-PCR techniques.     

水稻二化螟和三化螟基因组ddRADseq文库的构建

本文介绍了构建水稻二化螟和三化螟"双酶切限制性酶切位点关联DNA测序"(Double digest restrictionsite associated DNA sequencing,ddRADseq)文库的方法。利用安捷伦2100生物分析仪对4种单酶切及2种双酶切的酶切产物片段大小及分布范围进行分析,筛选出Mlu C I和Nla III两种限制性内切酶组合对螟虫基因组DNA进行酶切。酶切后的DNA片段两端连接上特定的P1、P2接头后,用Pippin Prep回收大小为285-435 bp的DNA片段。通过PCR扩增进行文库的富集并引入index序列。构建好的ddRADseq文库用琼脂糖凝胶电泳和生物分析仪进行质量检测。本方法所构建的文库DNA片段长度、分布和摩尔浓度能够达到Illumina平台测序的技术要求。本研究证实了利用Mlu C I和Nla III组合酶切构建水稻螟虫基因组ddRADseq文库的可行性,为在水稻螟虫中利用ddRADseq技术开展生物地理学、种群遗传学和系统发育重建等方面的研究奠定基础。     

Effects of insularity on genetic diversity within and among natural populations

We conducted a quantitative literature review of genetic diversity (GD) within and among populations in relation to categorical population size and isolation (together referred to as “insularity”). Using populations from within the same studies, we were able to control for between‐study variation in methodology, as well as demographic and life histories of focal species. Contrary to typical expectations, insularity had relatively minor effects on GD within and among populations, which points to the more important role of other factors in shaping evolutionary processes. Such effects of insularity were sometimes seen—particularly in study systems where GD was already high overall. That is, insularity influenced GD in a study system when GD was high even in non‐insular populations of the same study system—suggesting an important role for the “scope” of influences on GD. These conclusions were more robust for within population GD versus among population GD, although several biases might underlie this difference. Overall, our findings indicate that population‐level genetic assumptions need to be tested rather than assumed in nature, particularly for topics underlying current conservation management practices.     

Field evaluation of resistance of transgenic rice containing a synthetic cry1Ab gene from Bacillus thuringiensis Berliner to two stem borers

Two transgenic rice (Oryza sativa L.) lines, KMD1 and KMD2 at the R4 generation, transformed with a synthetic cry1Ab gene from Bacillus thuringiensis Berliner, were first evaluated for stem borer resistance in the field during the rice growing season of 1998 in two areas of Zhejiang Province, China. Both KMD1 and KMD2 were highly resistant to the stem borers Chilo suppressalis (Walker) and Scirpophaga incertulas (Walker), and were completely undamaged during the whole rice growing season. In contrast, damage to the plants of the untransformed parental control (Xiushui 11) was in the form of deadhearts or whiteheads. Under natural infestation by the C. suppressalis, the damage to control plants reached a peak of 88.7% of plants and 20.1% of tillers encountered with deadhearts. Under artificial and natural infestation of neonate striped stem borers at the vegetative stage and booting stage, 100% of plants and 25.6% of tillers, 78.9% of plants and 15.6% of productive tillers among artificially infested control plants were observed with the symptom of deadhearts and whiteheads, respectively. Damage to the control plants from artificial infestation by the S. incertulas reached a peak of 97.0% of plants and 22.9% of tillers damaged. The field research indicated that both KMD1 and KMD2 show great potential for protecting rice from attack by these two stem borers.