Phytoplasma transmission by in vitro graft inoculation as a basis for a preliminary screening method for resistance in fruit trees

In vitro grafting was tested as a technique for inoculating Prunus rootstock Prunus marianna GF 8-1 with European stone fruit yellows (ESFY) phytoplasmas and apple rootstock Malus pumila MM106 with apple proliferation (AP) phytoplasmas. In vitro shoot cultures of ESFY-infected Prunus marianna GF 8-1 and AP-infected Malus pumila MM106 were used as graft inoculum to transmit the phytoplasmas to the respective healthy rootstock. Phytoplasma transmission was assessed after a graft contact of 1, 2 or 3 months. Healthy autografts were used as controls to monitor parameters of in vitro grafting. Successful graft union formation ranged from 58 to 79% irrespective of the plant species and the sanitary state of the graft. Pathogen-specific polymerase chain reaction (PCR) was used to test the inoculated rootstocks for the presence of ESFY and AP phytoplasmas, respectively. The rate of ESFY phytoplasma transmission in successful Prunus -grafts increased from 69 to 94% with the time of contact. AP phytoplasma transmission to Malus occurred in 80 to 97% of successful grafts. To our knowledge this is the first report of phytoplasma transmission by grafting in vitro . The results provide a good basis for the establishment of a preliminary in vitro screening method for phytoplasma resistance in Prunus and Malus .     

浙江省广西越桔组织培养及遗传多样性分析

为了保育浙江省广西越桔,以凤阳山的广西越桔未成熟果实为材料,开展了未成熟种子培养、离体快繁体系建立、种群增强与回归和遗传多样性分析等研究。结果表明,接种至发育培养基(WPM+活性炭2.0 g·L^-1+谷氨酰胺0.4 g·L^-1+酶水解酪蛋白0.5 g·L^-1)后,85.19%未成熟种子发育完全,接种至萌发培养基(WPM+6-BA 1.0 mg·L^-1+GA₃ 1.5 mg·L^-1)后,30.43%发育完全的种子萌发,总萌发率为25.92%。最适增殖培养基为6号增殖培养基(WPM+ZT 0.2 mg·L^-1+NAA 0.5 mg·L^-1),增殖系数达106.67%,在生根培养基(WPM+IBA 1.0 mg·L^-1)上培养45 d后生根率为100%。种群增强与回归过程中植株长势良好,株高及株幅明显增加,部分植株与原生株及其他回归植株相比出现明显表型差异。遗传多样性分析结果显示,原生株与后代群体间的遗传相似系数在0.727 1~0.977 8之间,平均相似系数为0.841 0,极差为0.250 7。本研究成果可为浙江省广西越桔及相近处境珍稀植物的保育和利用提供理论与实践依据。     

鱼类工厂化循环水人工繁育设施装备应用研究进展

水产种业的高质量发展是推动养殖业发展的基础,现阶段中国鱼类人工繁育生产方式设施装备化程度低,产业大而不强,转型升级的需求迫在眉睫。该研究在文献调研整理的基础上,首先就循环水人工繁育设施装备在亲鱼产卵、鱼卵孵化、鱼苗培育等方面的研究和应用情况展开论述。相对于常规培育方式,循环水系统能够最大程度上构建出符合亲鱼交配和产卵的环境条件,养殖密度0.01～4.5 kg/m3,系统循环率9～76%/h,换水率0.7～3%/d;针对不同性质的鱼卵,介绍了国内外目前常用的孵化器主要有瓶式孵化罐、平列式孵化槽、漏斗式孵化器等,阐明了其适用对象、工作原理和主要性能表现;针对育苗和养殖系统构建需求的差异,综述了目前在循环水育苗设施装备应用研究中关注的重点和难点。其次,概述了中国鱼类人工繁育发展现状和问题,分析了循环水人工繁育技术的优势和面临的挑战。最后提出,鱼类工厂化循环水人工繁育具有较高的可行性和引领性,但是要实现产业化应用仍需要进一步开展基于品种对象的人工繁育环境构建及循环水处理、繁育过程鱼类对环境应激源的生物学响应、智能繁育装备等方面技术攻关和装备研发。     

甘蔗健康脱毒种苗生产技术方法研究初报

利用茎尖离体组织培养与热水处理相结合的技术方法,进行甘蔗脱毒健康种苗生产研究。试验结果表明:该方法生产的无病无毒的健康种苗,在出苗、分蘖、生长速度、株高、茎径、有效茎及产量等方面与对照比有显著差异,蔗茎增产幅度达26.65%～69.39%。     

打造日照绿茶品牌建设北方第一绿茶生产基地

本文介绍了日照市茶叶现状，提出了今后将其建设为北方第一绿茶生产基地的六点建议。     

沿海甜菜(Beta maritima L.)高频率丛生芽诱导和植株再生

选用沿海甜菜(Beta maritima L．)幼嫩花序项端为试验材料，建立了一个快速高效的离体培养体系。在附加1mg／L 6-BA的MS培养基上培养6周可诱导出丛生芽，诱导率达90％左右。丛生芽在适宜培养基上可快速扩增和长期继代培养。在附加1mg／L NAA的MS培养基上诱导小苗生根，生根率可达90％。再生植株经炼苗后移栽成活率为70％-80％。     

不同夜间温度处理对马铃薯试管薯及块茎形成相关基因表达的影响

为探究不同夜间温度处理对马铃薯试管薯形成的影响,以7个马铃薯品种或材料的试管苗为试材,在昼/夜温度分别为23℃/13℃、23℃/18℃和23℃/23℃条件下进行试管薯的诱导培养,调查记录试管薯初始结薯时间、结薯率和结薯方式变化,并分析不同夜间温度处理下试管薯初始形成植株及同期未形成试管薯植株参与块茎形成相关基因的表达量变化。结果表明,对于大多数基因型,与其他处理相比,23℃/13℃处理显著延长试管薯初始结薯时间,23℃/23℃处理降低结薯率,而23℃/18℃处理结薯较早且试管薯结薯率平均达84.3%,最利于试管薯形成。夜间温度可通过影响生物钟、糖代谢和光周期通路中StFKF1、StCO1和StSWEET1等相关基因的表达而调控块茎形成。     

Comparison of isolation and primary culture in vitro between human ovarian mural granulosa cells and cumulus cells

AIM To establish a suitable cell model for the study of ovarian function through comparing the isolation and primary culture effect of human ovarian mural granulosa cells （MGCs） and cumulus cells （CCs）. METHODS The follicular fluid of 16 patients who underwent assisted reproductive technique and their cumulus oocyte complexes （n=223） were collected. Density gradient centrifugation was used to isolate the MGCs and the methods of mechanical cutting plus enzyme hydrolysis were used to isolate the CCs. The cell counts and survival rates were analyzed by trypan blue staining and the expression of follicle stimulating hormone receptor （FSHR） was analyzed by flow cytometry to identify the purity. The expression of microtubule-associated protein 1 light chain 3 （LC3）, P62 and Bax at mRNA and protein levels was determined by qPCR and Western blot, respectively. RESULTS There had less isolation time, higher survival rate （P<0.05） and better tractility in vitro of CCs compared with MGCs. The results of flow cytometry showed that the FSHR expression of CCs and MGCs after isolation was （92.23±2.66）% and （81.33±6.57）%, respectively, with significant differences （P<0.05）. The mRNA level of LC3 in CCs was significantly lower than that in MGCs （P<0.01）, and the mRNA level of Bax was significantly higher than that in MGCs （P<0.05）. There was no significant difference in P62 mRNA expression between CCS and MGCs（P>0.05）. The difference of protein expressions of these molecules in the 2 kinds of cells were consistent with that in mRNA. CONCLUSION Mechanical cutting method plus enzyme hydrolysis is a simple way to isolate the CCs, with high purity and good cellular state in vitro, which can be used as a cell model for ovarian function research.     

农业综合开发投资绩效评价初探

本文用层次分析法对我国农业综合开发投资进行了分析,并根据投资效果对各省份进行了排序,进而对我国农业综合开发绩效进行了评价,并提出了几点结论性的建议。     

甜叶菊多倍体离体诱导技术体系的建立

以甜叶菊茎段为外植体,用不同浓度的秋水仙素在试管苗茎段再生植株过程中进行不同时间处理,探讨适合于甜叶菊离体多倍体诱导的最佳秋水仙素处理时间和浓度,初步建立甜叶菊离体诱导多倍体的技术体系。结果表明,秋水仙素对甜叶菊植株形成影响较大,秋水仙素处理的最佳剂量为0.1%处理3 d,再生植株总数高且多倍体比率达42.3%;同时,再生苗生根也受到了一定影响;多倍体苗叶片厚实、颜色较深、植株矮化,气孔增大,单位面积气孔数较少;多倍体细胞核体积增大,染色体数加倍。