鸡皮刺螨实验室饲养与繁殖技术研究进展

鸡皮刺螨是一种吸血性外寄生虫,会引起鸡消瘦、贫血,并导致产蛋下降,造成巨大经济损失。为给科研提供可靠虫源,并对杀螨药物、疫苗以及其他防控手段进行效果评价,国内外学者进行了鸡皮刺螨实验室的人工饲养与繁殖技术研究。文章综述了国内外鸡皮刺螨实验室饲养与繁殖技术的研究进展,将其总结归纳为使用动物饲养繁殖技术与离体饲养繁殖技术两类,同时指出了各项技术在应用过程中存在的问题,为该技术的进一步完善与发展提供相关参考。     

超表达OsPR1A增强了Xa21介导的水稻对白叶枯病的抗性反应

【背景】前期研究发现,水稻病程相关蛋白质OsPR1A的表达受上游抗病基因Xa21调控,接菌后早期启动Xa21介导的OsPR1A较高水平表达对水稻抵抗白叶枯病菌至关重要。同时OsPR1A也受到水稻白叶枯病菌（Xanthomonas oryzae pv. oryzae,Xoo）的诱导表达。对于OsPR1A的研究绝大部分是作为抗性反应发生的标志基因佐证其他基因或途径在抗性中的作用,缺乏直接的证据证实OsPR1A本身的生物学功能。【目的】通过获得OsPR1a-OX超表达转基因植株,调查其表型及农艺性状,并明确OsPR1A蛋白质表达与抗性的关系,为鉴定OsPR1A功能提供依据。【方法】通过农杆菌介导法,将构建的OsPR1a-OX转化载体转入到水稻受体4021中,利用PCR和免疫印迹（western blot,WB）技术分别在基因水平和蛋白质水平上筛选并鉴定OsPR1A超表达阳性纯合株系。在成熟期,调查OsPR1A超表达转基因植株的表型及农艺性状（株高、穗长、分蘖数、结实率和籽粒大小等）。在31℃条件下,将生长2周的水稻幼苗TP309、4021和OsPR1A超表达转基因植株接种水稻白叶枯病菌,并在接菌0、2、4、6、8、10和12 d时测量病斑长度。在接菌0、4和6 d时,收集TP309、4021和OsPR1A超表达转基因植株的水稻叶片,提取蛋白质,利用WB技术检测OsPR1A的表达特征。【结果】构建了OsPR1a-OX转化载体,并转入到受体4021中,筛选并鉴定到2个OsPR1A超表达转基因纯合株系（#704和#709）。调查了OsPR1A超表达转基因植株在成熟期的表型及农艺性状,与对照4021相比,#704和#709的株高较矮、穗长较短、分蘖数减少、结实率降低,但籽粒稍大,可能与结实率低有关。在31℃条件下,OsPR1A超表达转基因植株的病斑长度与对照4021相比明显缩短,结果具有显著性差异（P<0.05）。在接菌0、4和6 d的材料中,超表达转基因植株#704和#709中OsPR1A始终有较高水平的表达丰度,从而提高了对白叶枯病菌的抗性。【结论】采用农杆菌介导法,获得OsPR1A超表达转基因植株;超表达OsPR1A影响到水稻的正常发育过程;超表达OsPR1A后增强了Xa21介导的水稻对白叶枯病的抗性。     

草地贪夜蛾对小麦和玉米的产卵选择性及其种群生命表

为明确草地贪夜蛾对小麦的产卵选择性及其是否对小麦安全生产构成威胁,本研究以玉米和小麦作为测试寄主,比较分析了草地贪夜蛾对两种作物不同部位的产卵选择性,并利用两性生命表方法研究了取食小麦、玉米对其生命参数的影响。结果表明:草地贪夜蛾更喜欢在玉米上产卵,其在玉米、小麦叶片、玉米和小麦茎秆上的产卵量存在显著差异(df=102,F=15.593,P<0.05),以玉米叶片背面卵块数量(7.11±1.55)块/笼最高;草地贪夜蛾取食小麦可以完成生活史,但幼虫存活率、化蛹率、羽化率和世代存活率低于取食玉米。取食玉米的幼虫发育历期为(16.31±0.15)d,显著高于取食小麦的(14.66±0.12)d,蛹期、蛹重、产卵前期、成虫寿命和世代周期无显著差异。取食小麦羽化出的雌虫寿命、平均单雌产卵量显著高于取食玉米,分别为(16.39±0.40)d、(976.31±57.21)粒和(14.64±0.32)d、(831.57±30.55)粒。生命表参数显示取食玉米的净增殖率为363.14,显著高于小麦的258.63,但内禀增长率、周限增长率和平均世代周期无显著差异。研究结果为草地贪夜蛾在小麦上的预测预报和有效防控提供了基础数据。     

The M43 domain-containing metalloprotease RcMEP1 in Rhizoctonia cerealis is a pathogenicity factor during the fungus infection to wheat

Wheat(Triticum aestivum L.) is an important staple crop for global human. The necrotrophic fungus Rhizoctonia cerealis is the causal pathogen of sharp eyespot, a devastating disease of wheat. Herein, we identified RcMEP1, a zinc metalloproteaseencoding gene from R. cerealis genomic sequences, and characterized its pathogenesis function. RcMEP1 expressed at markedly-high levels during R. cerealis infection process to wheat. The predicted protein RcMEP1 comprises of 287 amino acid residues and contains a signal peptide and a M43 metalloprotease domain harboring the active site motif(HEVGHWLGLYH). The assays of Agrobacterium tumefaciens-mediated transient expression in Nicotiana benthamiana leaves indicated that RcMEP1 is an apoplastic elicitor of cell death, and that the predicted signal peptide functions and is required for secretion and cell death-induction. The purified RcMEP1 protein and its M43 domain peptide were individually able to induce plant cell death and H2 O2 accumulation, and to inhibit expression of host chitinases when infiltrated into wheat and N. benthamiana leaves, while the M43 domain-deleting peptide and negative control lacked the capacity. Moreover, compared with the control pretreatment, the purified RcMEP1 protein or its M43-domain peptide resulted in enhanced pathogenesis in the inoculated wheat, whereas the M43 domain-deleting peptide failed. These results suggest that RcMEP1 acted as an important pathogenicity factor during R. cerealis infection to wheat and that its signal peptide and M43 domain are required for the secretion and pathogenesis of RcMEP1. This study provides insights into pathogenesis role of M43 domain-containing metalloproteases during R. cerealis infection to wheat.     

Effect of dietary calcium or phosphorus deficiency on bone development and related calcium or phosphorus metabolic utilization parameters of broilers from 22 to 42 days of age

This experiment was conducted to investigate the effect of dietary calcium (Ca) or phosphorus (P) deficiency on bone development and related Ca or P metabolic utilization parameters of broilers from 22 to 42 days of age based on our previous study, which indicated that dietary Ca or P deficiency impaired the bone development by regulating related Ca or P metabolic utilization parameters of broilers from 1 to 21 days of age. A total of 504 one-day-old Arbor Acres male broilers were randomly assigned to 1 of 4 treatments with 7 replicates in a completely randomized design, and fed the normal control and Ca- or P-deficient diets from 1 to 21 days of age. At 22 days of age, the broilers were further fed the normal control diet (0.90% Ca+0.35% non-phytate P (NPP)), the P-deficient diet (0.90% Ca+0.18% NPP), the Ca-deficient diet (0.30% Ca+0.35% NPP) or the Ca and P-deficient diet (0.30% Ca+0.18% NPP), respectively. The results showed that dietary Ca or P deficiency decreased (P<0.05) tibia bone mineral density (BMD), bone breaking strength (BBS), ash content, tibia ash Ca content and serum P content on days 28 and 42, but increased (P<0.05) tibia alkaline phosphatase (ALP) activity of broilers on day 42 compared with the control group. Furthermore, the broilers fed the P-deficient diet had the lowest (P<0.05) tibia BMD, BBS, ash content, serum P content and the highest (P<0.05) serum Ca content on day 28 compared with those fed the Ca-deficient or Ca and P-deficient diets. The results from the present study indicated that the bone development and related Ca or P metabolic utilization parameters of broilers were the most sensitive to dietary P deficiency, followed by dietary Ca deficiency or Ca and P-deficiency; dietary Ca or P deficiency impaired the bone development possibly by regulating serum Ca and P contents as well as tibia Ca content and ALP activity of broilers from 22 to 42 days of age.     

苹果花脸病田间病情发展及ASSVd在组培条件下的传播

为了明确苹果锈果类病毒（apple scar skin viroid，ASSVd）引起的苹果花脸病在田间的病情发展情况以及ASSVd在组培条件下的传播方式，2012—2015年对河北省顺平南神南村3个果园进行持续调查和检测。结果显示，果园A、B、C的显症率分别从3.00%、19.35%和10.00%上升至10.00%、34.41%和22.00%，带毒率分别从11.00%、20.43%和14.00%上升至23.00%、37.63%和30.00%，带毒率和显症率都逐年增加，而且带毒率大于显症率，即有些带毒果树不显症。发病果树田间分布有明显的发病中心。通过高通量测序发现显症果树ASSVd vsiRNA reads数是带毒未显症reads数的4.08倍，同时实时荧光定量PCR检测发现显症树ASSVd病毒积累量显著高于带毒未显症树。以携带ASSVd的组培苗和无毒组培苗为试材，利用RT-PCR检测明确了ASSVd可通过伤口沾染带毒汁液、组培剪污染、含毒培养基、根系接触进行传播，其中根系接触传染率较高。     

华北落叶松不同代际人工林土壤养分及细菌群落变化特征

目的探究不同代际华北落叶松人工林土壤养分及细菌变化有助于了解地力维持机制。为华北落叶松人工林可持续经营提供参考。方法本研究以塞罕坝地区华北落叶松不同代际人工林（一代林（17和37年生）、二代林（14年生））为研究对象，分析了不同代际林及不同土层土壤养分、土壤细菌群落组成和多样性的变异规律及土壤养分与细菌群落的耦合关系。结果（1）在0 ~ 10 cm和10 ~ 20 cm土层，一代林（17年生）和二代林（14年生）的速效钾和有效磷显著高于一代林（37年生）（P < 0.05），但一代林（17年生）和二代林（14年生）间差异不明显（P > 0.05）。随着土壤深度增加，各代际林土壤养分均呈现降低趋势。（2）在门水平上，3个林分的优势菌群均为变形菌门、放线菌门、疣微菌门和酸杆菌门。随着土层深度的增加，3个林分的变形菌门和放线菌门呈迅速下降趋势。（3）一代林（37年生）的ACE和Chao1细菌丰富度指数在3个代际林中最低，且在0 ~ 10 cm和20 ~ 30 cm土层代际林间差异显著（P < 0.05）。在3个土层深度，土壤细菌多样性指数在一代林（17年生）和二代林（14年生）中相对较高，而在一代林（37年生）中相对较低。（4）相关分析表明，速效钾与变形菌门和放线菌门显著正相关，而与硝化螺旋菌门显著负相关（P < 0.05）。随着有效磷含量的增加，变形菌门数量呈显著增加趋势（P < 0.05）。结论华北落叶松二代林在早期发育阶段不存在地力衰退问题，随着林龄的增加，应加强速效钾、有效磷和碱解氮的调控，并重视特定细菌群落的变化以维持土壤肥力。      

Screening of antagonistic Trichoderma strains and their application for controlling stalk rot in maize

Maize is one of the major crops in China, but maize stalk rot occurs nationwide and has become one of the major challenges in maize production in China. In order to find an environment-friendly and feasible technology to control this disease, a Trichoderma-based biocontrol agent was selected. Forty-eight strains with various inhibition activities to Fusarium graminearum, and Fusarium verticillioides were tested. A group of Trichoderma strains(DLY31, SG3403, DLY1303 and GDFS1009) were found to provide an inhibition rate to pathogen growth in vitro of over 70%. These strains also prevented pathogen infection over 65% and promoted the maize seedling growth for the main root in vivo by over 50%. Due to its advantage in antifungal activity against pathogens and promotion activity to maize, Trichoderma asperellum GDSF1009 was selected as the most promising strain of the biocontrol agent in the Trichoderma spectrum. Pot experiments showed that the Trichoderma agent at 2–3 g/pot could achieve the best control of seedling stalk rot and promotion of maize seedling growth. In the field experiments, 8–10 g/hole was able to achieve over 65% control to stalk rot, and yield increased by 2–11%. In the case of natural morbidity, the control efficiency ranged from 27.23 to 48.84%, and the rate of yield increase reached 11.70%, with a dosage of Trichoderma granules at 75 kg ha~(-1). Based on these results, we concluded that the Trichoderma agent is a promising biocontrol approach to stalk rot in maize.     

MYB转录因子参与植物非生物胁迫响应与植物激素应答的研究进展

基因的时空表达受转录因子的精确调控。植物在面对不利环境因素比如高温、低温、干旱、盐碱等胁迫时其细胞生理生化会迅速地从“舒适”状态转变进入“胁迫响应”状态。这种快速响应的状态转变依赖于植物对胁迫信号的感知及传递、激素通路(脱落酸、茉莉酸等)的激发、转录因子的活化等复杂的过程;最终植物通过胁迫相关基因的表达、次生代谢转变、抗氧化物质的积累等实现胁迫条件下细胞内环境的再平衡从而获得生存。植物MYB(v-MYB avian myeloblastosis viral oncogene homolog)转录因子就是上述转变中的重要参与者。本文介绍了植物MYB转录因子的结构特征、分类,综述了近些年来MYB转录因子与非生物胁迫,以及植物激素应答过程相关的研究进展。