光枝勾儿茶内生真菌多样性及抑菌活性研究

为探索贵州苗药光枝勾儿茶内生真菌类群特征、分布部位及其抑菌活性,该研究采用传统方法对贵州省贵阳市和黔西市光枝勾儿茶内生真菌进行分离,并基于分子生物学及统计学对其分类地位进行鉴定及多样性评价,最后通过微量肉汤倍比稀释法筛选具有抑菌活性的菌株。结果表明:(1)从光枝勾儿茶中分离到191 株内生真菌,隶属于3 个门5 个纲10 个目15 个科19 个属,优势属为叶点霉属(Phyllosticta)、间座壳属(Diaporthe)、葡萄座腔菌属(Botryosphaeria)和刺盘孢属(Colletotrichum)。(2)黔西光枝勾儿茶内生真菌香农-维纳多样性指数(H''_Q=2.112)较贵阳(H''_G=1.801)高,索伦森相似性指数Cs_G-Q为0.923,不同组织香农-维纳多样性指数为茎(H''_S=2.004)>根(H''_R=1.764)>叶(H''_L=1.654)>果实(H''_F=1.473),茎和叶内生真菌的索伦森相似性最高(Cs_S-L=0.667)。(3)筛选出的21株内生真菌对供试菌大肠杆菌(Escherichia coli)、金黄色葡萄球菌(Staphylococcus aureus)和沙门氏菌(Salmonella enterica)具有抑菌效果,其中Diaporthe sp. QX4G6对大肠杆菌、金黄色葡萄球菌和沙门氏菌的最小抑菌浓度分别为12.5、6.25、12.5 mg·mL^-1,最小杀菌浓度分别为12.5、6.25、12.5 mg·mL^-1。以上研究结果揭示了光枝勾儿茶蕴藏丰富的内生真菌资源,不同地区及组织内生真菌类群组成有差异,多个分离菌株具有抗菌活性,为光枝勾儿茶内生真菌天然抗菌药物或药源研发奠定了基础。     

Effect of (E4, Z6)-4,6-Hexadecadienal in attraction of Stathmopoda masinissa with its pheromone components of Korean population

Timely insecticidal application for Stathmopoda masinissa Meyrick (Lepidoptera: Stathmopodidae), is important, for reducing damage to persimmon (Diospyros kaki Thunb.), an important tree fruit cultivated in Korea. In this regard, the early and precise detection of adult S. masinissa is desirable. In this study, we report the effect of (E4,Z6)-4,6-hexadecadienal (E4,Z6-16Ald) with sex pheromone components in attracting S. masinissa males. The sex pheromone of S. masinissa in the Korean population comprised two components, (E4,Z6)-4,6-hexadecadienyl acetate (E4,Z6-16Ac) and (E4,Z6)-4,6-hexadecadienol (E4,Z6-16OH). It was shown that the E4,Z6-16Ald acts as a synergist of E4,Z6-16Ac for attracting S. masinissa in the Japanese population. To test whether E4,Z6-16Ald could be used as an attractant in the Korean population, the E4,Z6-16Ald with the two pheromone components was evaluated in attracting S. masinissa males. Electroantennography (EAG) assays were performed to determine the antennal responses of S. masinissa males to the two pheromone components and E4,Z6-16Ald tested. A field attraction test with a combination of pheromones and E4,Z6-16Ald was carried out for 3 years in three different regions in Korea. E4,Z6-16Ald elicited as high a response as the two pheromone components. A mixture of the two pheromone components and E4,Z6-16Ald and a mixture of E4,Z6-16Ac and E4,Z6-16Ald attracted more S. masinissa males than a mixture of E4,Z6-16Ac and E4,Z6-16OH, the pheromone of Korean population. This new pheromone lure formulation with E4,Z6-16Ald is expected to contribute to the precise detection of S. masinissa by luring males to pheromone-baited traps.     

Stage specific feeding attributes and mobilization of nutrients in Mexican beetle,Zygogramma bicolorata Pallister on Parthenium hysterophorus L.

Parthenium hysterophorus L. (Asteraceae) is a toxic weed of agricultural farms, pastures and wastelands with a pan-tropical distribution. The weed causes a reduction in crop production of agricultural fields and severe health problems in humans. The Mexican beetle, Zygogramma bicolorata Pallister (Coleoptera: Chrysomelidae) is among the most promising candidates for the weed biocontrol. However, no previous study has evaluated assimilation of nutrients in this beetle. In the present study, feeding attributes and assimilation of nutrients by larvae of Z. bicolorata were assessed on the weed. Results revealed that the first larval instars of Z. bicolorata exhibited lowest consumption rates, and accumulated minimum concentrations of glucose, proteins and triglycerides in their body. They showed compensatory feeding, and displayed highest food utilization efficiencies and developmental rates. In contrast, the fourth larval instars exhibited higher food consumption rates and conversion efficiencies, but displayed lowest developmental rates. Accumulation of food reserves was also recorded maximum for the fourth larval instars. Overall, the findings may aid current mass-rearing efforts for Z. bicolorata in laboratories, but field trials are still needed to strengthen the present findings.     

The abundance and diversity of gut bacteria of rice leaffolder Cnaphalocrocis medinalis (Guenée) across life stages

The bacterial community living in the insect gut may play an important role in nutrition, immunity and protection, detoxification of toxins, and inter- and intra-specific communication. Rice leaffolder Cnaphalocrocis medinalis (Guenée) (Lepidoptera: Crambidae) is a notorious pest in rice, and the diversity of the gut bacteria of C. medinalis across life stages are not well understood. Here, the diversity and abundance of the gut bacterial community in C. medinalis through life stages were investigated using Illumina Miseq technology. A total of 22 bacterial phyla, 42 classes, 100 orders, 179 families, 350 genera and 395 species were identified across the different life stages of C. medinalis. Proteobacteria and Firmicutes phyla were the dominant bacterial taxa. Members of the genera Enterococcus, unclassified Enterobacteriaceae, Wolbachia, Acinetobacter, Stenotrophomonas, Microbacterium, Bacillus, Corynebacterium, Lampropedia, and Sphingobacterium were found at all life stages. Enterococcus and unclassified Enterobacteriaceae occupied higher relative abundance among bacteria community in the 2nd to 5th instar larvae, pupae and adults. The structure of bacterial community differed across the life stages of C. medinalis. Our findings will enrich the understanding of gut bacteria in C. medinalis, and will provide foundation and assistance for the development of novel pest management strategies through utilization of microbiota.     

Changes in spatial and temporal distributions of Monochamus beetles along the fire severity in burned Pinus densiflora forests

Wildfire is an important disturbance factor in forest ecosystem and could affect the distribution of insects. Because Monochamus beetles are known vector insects of pine wood nematodes in Korea and burnt pine trees can be used as oviposition habitats for Monochamus beetles, we investigated changes in the spatial (among fire severity classes) and temporal (between years) distributions of M. alternatus and M. saltuarius. From late May 2017 to early October 2018, four multi-funnel traps with pheromone lure were installed in each fire severity class (unburned, light, light-moderate edge, moderate-severe edge, and severe). All traps were replaced fortnightly from May to October for two years. The spatial distribution of the two Monochamus beetles was significantly affected by fire severity. In particular, M. alternatus showed higher catches in the severely burned sites and responded positively to the fire. A similar number of M. saltuarius individuals were observed between 2017 and 2018, while more M. alternatus were caught in 2018 compared to 2017. Consequently, it was confirmed that the dead wood of P. densiflora caused by the wildfire had a great influence on the density of insects, especially M. alternatus.     

Genetic diversity and geographic distribution of Bemisia tabaci species complex in Nepal

Bemisia tabaci species complex comprises at least 44 cryptic species worldwide. Here, we analyze the nucleotide sequences of mitochondrial cytochrome oxidase subunit 1 (COI) gene obtained from 76 samples of B. tabaci collected from 23 districts in Nepal. This is the first genetic and geographic study of B. tabaci species complex in Nepal. Our phylogenetic study identified the presence of three cryptic species—Asia I, Asia II 1, and Asia II 5—with high interspecific but low intraspecific variations. Among the three cryptic species, Asia II 5 was the most prevalent in Nepal, constituting 64.47% of all the sequenced samples. Based on haplotype network analysis of COI sequences, Asia II 1 was more genetically diversified than the other two cryptic species. Our results provided useful information on the genetic diversity and geographic distribution of B. tabaci in Nepal, which help monitor population changes of B. tabaci at cryptic species level and develop sustainable management strategies for its control.     

An effective entomopathogenic fungus Metarhizium anisopliae for the simultaneous control of Aedes albopictus and Culex pipiens mosquito adults

The purpose of this study was to select the most effective Metarhizium anisopliae isolates for the simultaneous control of Aedes albopictus and Culex pipiens mosquito adults. The selection was performed using 65 isolates of M. anisopliae, an entomopathogenic fungus isolated from Korean soils. The selection process was carried out through evaluations of conidial thermotolerance, UV-B tolerance, pathogenicity and virulence against adults of two mosquito species and conidial production on grain medium. The thermotolerance of 65 M. anisopliae isolates ranged from 0% to 99%, and their UV-B tolerance ranged from 7.1% to 94.8%. Among them, 30 isolates with high thermotolerance were selected for evaluation of their pathogenicity and virulence against adults of the two mosquito species. Metarhizium anisopliae isolates were more susceptible to Cx. pipiens adults than Ae. albopictus adults because 11 isolates were pathogenic to Ae. albopictus and 21 isolates were pathogenic to Cx. pipiens. Virulence and conidial production were evaluated using 6 isolates with similar pathogenicity against the two species of mosquito. As a result, the accumulated mortality was 74–100% for Ae. albopictus and 78–100% for Cx. pipiens. LT₅₀ values ranged from 2.3 to 5.2 days for Ae. albopictus and from 3.2 − 4.7 days for Cx. pipiens. Comparison of conidial production using three kinds of grain media showed various results according to isolate and medium. Based on the above results, M. anisopliae CN6S1W1 was finally selected as the most effective fungal isolate for the simultaneous control of Ae. albopictus and Cx. pipiens mosquito adults.     

Artificial radiofrequency driven life-table parameters of perilla seed bugs (Nysius sp.) (Heteroptera: Lygaeidae)

The genus Nysius (Hemiptera: Lygaeidae) are polyphagous pests of various plants that damage seeds, foliage, and flowers. Here, we studied the life-table parameters and adult performances of Nysius plebeius Distant and Nysius hidakai Nakatani after exposure to five radiofrequencies: 0 (control), 5, 10, 20, and 30 kHz in infested perilla seeds. In addition, we measured the follow on effects of treatment for four generations (Parent, F1-, F2-, and F3-generation), for both Nysius species. Radiofrequency application significantly affected the life-table parameters of both Nysius species. Radiofrequency exposures affected bug oviposition, developmental period, adult weight, and longevity. Exposure also negatively affected the fecundity of subsequent generations; total developmental period from egg to adult emergence was longer while adult longevity was shorter in the radiofrequency treatments than in the untreated control. Low radiofrequency levels had the greatest effect on the life-table parameters of Nysius species, with the lowest rate of adult emergence and the shortest adult longevity at 5 kHz. The potential of using Nysius bug susceptibilities to radiofrequency as an alternative to chemical treatment is discussed.     

茂兰喀斯特区68种典型植物叶片化学计量特征

研究茂兰喀斯特区不同功能(类)群植物叶片的养分含量及化学计量特征,揭示其在时间和空间尺度上的变化规律,阐明碳(C)、氮(N)、磷(P)、钾(K)等养分含量与C∶N∶P间的相互关系,探讨N∶P对该区域植物生长的指示作用,以期能够更深入的了解其养分利用状况及适生性,为喀斯特森林的稳定性及维持机制提供理论依据。以茂兰喀斯特区68种典型植物为研究对象,分别测定不同生长阶段植物叶片的C、N、P和K含量,并计算其化学计量比。结果表明:研究区68种植物分属40科62属;其叶片C、N、P和K含量的几何平均值分别为445.87 g/kg、17.32 g/kg、1.35 g/kg和9.86 g/kg,C∶N的算术平均值为26.93,C∶P、C∶K、N∶P、N∶K和P∶K的几何平均值分别为330.93、45.22、12.85、1.76和0.137;C与N呈极显著负相关,N与P、K以及P与K均呈极显著正相关,N与C∶P和C∶K、P与C∶N、C∶K和N∶K以及K与C∶N、C∶P和N∶P均呈极显著负相关,且它们之间均具有二次函数、指数函数或幂函数的非线性耦合关系;从变异程度来看,C含量为弱变异,N、P、K含量及各元素的化学计量比则均属中等变异或强变异。从植物不同生活型来比较,各生长阶段的C含量均表现为灌木乔木草本,N、P和K含量均为草本灌木乔木,各元素的化学计量比则均为乔木灌木草本。从植物不同系统发育来分析,各生长阶段蕨类植物的N、P、K含量均要高于种子植物,而各元素的化学计量比则正好相反。从不同生长阶段来看,各功能(类)群植物生长期(或生长盛期)的养分含量均要高于落叶期(或生长末期);乔木、灌木和草本等不同生活型植物落叶期的C∶P、C∶K、N∶P和N∶K均要高于生长期;蕨类植物各元素的化学计量比不同生长阶段间差异都不显著;而种子植物的C∶P、C∶K、N∶P和N∶K则均表现为落叶期生长期。对比我国其他地区及全国和全球尺度上的研究结果,该区域植物的生长发育易受N和P素的双重限制,但又因功能(类)群及生长阶段的不同其受限的养分元素也存在一定差异,体现了对高度异质的喀斯特生境不同的适应策略;而植物体内较高的K含量则可能是提高其自身抗性、适应恶劣环境的重要因素。减少人为干扰、加之适当的保护,在植物生长期配以适量的N素添加,有利于其更好的生长发育,有助于提高喀斯特森林生态系统的稳定性和抗干扰性。研究结果揭示了喀斯特森林植物的适生机制,对喀斯特森林的保护具有重要的指导意义。     

Over-Expression of Masson Pine PmPT1 Gene in Transgenic Tobacco Confers Tolerance Enhancement to Pi Deficiency by Ameliorating P Level and the Antioxidants

Plant Molecular Biology Reporter - Previously, we cloned the full sequence of masson pine (Pinus massoniana) phosphate transporter gene (PmPT1) from a phosphorus (Pi) deficiency tolerant strain. To...