小麦TaNAC基因基于可变剪切和microRNA的转录后调控分析

【目的】以条锈菌和白粉菌胁迫的普通小麦（Triticum aestivum L.）为研究对象,分析由可变剪切（alternative splicing,AS）形成的TaNAC结构变异转录本,同时分析TaNAC基因的microRNA调控位点,为进一步解析TaNAC基因通过转录后调控参与小麦响应真菌胁迫奠定基础。【方法】普通小麦兼抗种质N9134在被白粉菌和条锈菌分别侵染后,各8个时间点取样并混合,然后从混合样本中克隆得到大量TaNAC转录本。参考中国春小麦基因组注释信息（IWGSC RefSeq v1.1）进行比对,选择由可变剪切形成的TaNAC序列结构变异转录本,分析它们的序列结构特征。利用生物信息学软件和在线工具,对这些TaNAC结构变异转录本编码产物的功能结构域、高级结构、理化性质、亚细胞定位等特征和变异情况进行比对分析。同时,利用洋葱表皮细胞瞬时表达系统验证其中1对TaNAC结构变异转录本的亚细胞定位预测结果,并选取5组TaNAC基因的可变剪切序列结构变异转录本进行酵母转录自激活试验,研究序列结构变异对TaNAC基因转录调控活性的影响。此外,利用miRBase数据库收录的小麦中已报道的miRNAs和TaNAC基因,进行靶基因预测分析,建立小麦TaNAC家族成员与miRNAs的靶向关系。【结果】以条锈菌和白粉菌侵染的普通小麦兼抗种质N9134为材料,克隆得到的TaNAC转录本中的35条序列结构变异转录本由13个TaNAC基因可变剪切形成。通过分析发现,同一TaNAC基因由可变剪切形成的不同结构变异转录本的核酸序列结构存在差异,而且其对应编码产物的功能结构域、高级结构、理化性质和亚细胞定位等方面均会存在差异,同时也会表现出不同的转录调控活性;不同TaNAC基因的可变剪切方式存在差异,而且它们的结构变异转录本及其编码产物在结构特征、理化性质和转录调控活性等方面也均呈现出多样性的特征。通过分析TaNAC基因与其在编码序列区域的靶标tae-miRNAs,发现具有可变剪切转录本的TaNAC基因与tae-miRNA的结合位点均在其非可变剪切区域。【结论】TaNAC基因可以通过可变剪切这种转录后调控方式参与小麦对真菌胁迫的响应;同时发现,调控TaNAC基因的tae-miRNA可以独立于可变剪切这种转录后调控方式行使功能。     

促进堆肥二次发酵真菌在堆肥中的应用效果

【目的】二次发酵是实现鸡粪等养殖废弃物功能价值提高的关键阶段,本研究旨在将前期筛选得到的真菌应用于鸡粪堆肥的二次发酵阶段,探讨其促进二次发酵的效果并对其进行鉴定,为今后鸡粪的高值化与资源化利用提供理论依据和支撑。【方法】以鸡粪一次发酵产物为原料,添加实验室前期筛选的2株真菌（分别为FCM1和FCM3）进行二次发酵试验。共设置3个处理,分别为对照CK：原料中加入1%（体积L∶干物质量kg）无菌液体培养基;F1：原料中加入1% FCM1菌液;F3：原料中加入1% FCM3菌液。在二次发酵的不同时期,分析测定堆体温度、pH、电导率（EC）、铵态氮、硝态氮、种子发芽指数以及腐殖酸等指标。【结果】与对照相比,添加真菌主要促进了二次发酵阶段快速升温和腐殖化过程快速进行。与未接菌处理相比,真菌FCM3使堆料提前8 d进入升温期,真菌FCM1效果不佳,且在一定程度上延后了鸡粪堆肥的二次发酵。F3处理高温阶段（>50℃）持续了13 d,堆料的GI比对照处理提前25 d达到80%,且腐殖化程度较高（PHA=74.58%,PHA：胡敏酸/总腐殖酸）。相关分析表明,GI与pH、PHA显著正相关,与EC显著负相关。经形态和ITS rDNA测序等方法鉴定,FCM3为链格孢菌。【结论】真菌FCM3能够在一定程度上促进鸡粪堆肥二次发酵的进行,提高堆料的腐殖化程度。     

人工湿地砷污染去除研究进展

砷在环境中普遍存在,因其高毒性,世界卫生组织将砷污染列为重大公共卫生问题。因此,科学家们在不断寻求科学有效的砷污染去除方法。人工湿地砷污染去除系统自上世纪七十年代推出以来,因其环境友好、成本低廉的特性得到蓬勃发展,快速成为绿色环保修复技术之一。人工湿地被认为是理想的砷富集去除场所,其对砷污染的去除方式主要通过植物和微生物相互作用及腐殖酸强化修复来实现。本文通过砷污染治理历史及技术发展、人工湿地的兴起及砷在湿地生态系统中的赋存状态以及人工湿地生态系统砷污染去除三个方面综述了人工湿地砷污染去除的研究进展。通过大量文献综述分析,我们认为在人工湿地砷污染去除的后续研究工作中,应着重研究砷在湿地生态系统中的扩散迁移及演变规律,构建湿地生态系统沉水-浮水-挺水植物立体结构,并不断探索植物-微生物及腐殖酸强化修复对砷污染的协同阻滞及界面作用。     

耦合视角下生产性服务业与智慧农业发展的研究——基于美国的经验与启示

为了推进农业生产性服务业与智慧农业的耦合,进而为我国小农户衔接智慧农业提供桥梁。本文选取美国为典型案例,分析美国生产性服务业与智慧农业耦合发展的模式和措施。采用比较研究法,归纳中美生产性服务业与智慧农业耦合发展的异同。研究表明,美国生产性服务业与智慧农业发展具有服务耦合、产业耦合和利益耦合三种模式,美国为推进其耦合发展而采取了立法保障、信息化建设、农业大数据利用和高素质人才培养等一系列措施。中美两国在智慧农业的高门槛、农业人口流失与老龄化问题、突发性公共卫生危机等方面存在共同点。但与美国相比,中国的生产性服务业与智慧农业之间的关系尚处于并行分离的状态,存在耦合层次较低,创新驱动力不足等问题。因此,为推动生产性服务业与智慧农业耦合发展,提出健全相关配套支持机制、加强大数据平台建设和信息化技术研发、完善农业产业链条和加强专业性人才的培养力度等对策建议。     

安徽省稻瘟病菌生理小种鉴定分析

【目的】探究安徽省稻瘟病菌优势种群和优势生理小种分布,分析优势生理小种的致病力,为稻瘟病抗性育种和品种推广布局提供参考。【方法】利用7个水稻鉴别品种,采用苗期人工喷雾接种法对2019—2020年收集的安徽省5个稻作区21个市(县、区)的稻瘟病菌进行鉴定,利用不同优势生理小种对775份水稻资源进行苗瘟抗性鉴定,分析其致病力,筛选抗稻瘟病优异水稻资源。【结果】共鉴定出7群25个稻瘟病菌生理小种,优势种群为ZB群,出现频率为58.58%,其次为ZC群,出现频率为22.62%;优势生理小种为ZB13、ZB15和ZC15,出现频率分别为32.43%、17.98%和17.98%。沿淮淮北单季稻作区凤台县优势生理小种为ZB15、ZC15和ZB13,怀远县为ZB13和ZB9,谢家集区为ZB13和ZG1;江淮丘陵单、双季稻过渡区全椒县优势生理小种为ZB15,寿县为ZB13、ZC15和ZB15,肥西县为ZB13和ZB15;沿江双、单季稻作区青阳县优势生理小种为ZC15,无为市为ZB13和ZG1,宣州区为ZB13、ZB15和ZC15,当涂县为ZB13、ZC13和ZC15,贵池区为ZB15和ZC15,南陵县为ZC15、ZB13和ZB15,怀宁县为ZB13;大别山地单、双季稻作区金寨县优势生理小种为ZB13和ZG1,潜山市为ZB13和ZB15,岳西县为ZB13和ZH1;皖南山地单、双季稻作区石台县优势生理小种为ZB15、ZB13和ZC15,休宁县为ZC15、ZG1、ZB15和ZH1,宁国市为ZB13和ZG1,黄山区为ZB13、ZC13和ZC15,歙县为ZC15。优势生理小种致病力分析结果显示,休宁县和金寨县ZB13的致病力分别为59.68%和62.06%,平均为60.87%;休宁县和潜山县ZB15的致病力分别为61.16%和64.84%,平均为63.00%;休宁县和南陵县ZC15的致病力分别为71.29%和66.26%,平均为68.78%。通过筛选获得10份抗苗瘟优异种质资源。【结论】安徽省稻瘟病菌优势种群为ZB和ZC群,优势生理小种为ZB13、ZB15和ZC15。明确了安徽省不同稻作区生理小种的结构分布。     

生姜主栽品种枯萎病抗性评价及抗病性鉴定指标筛选

【目的】筛选抗枯萎病的生姜品种,建立抗病评价体系,为生姜枯萎病抗病品种选育及抗病机理研究提供理论参考。【方法】选取5个生姜主产区的主栽品种,采用室内人工接种法测定枯萎病菌(腐皮镰刀菌)侵染对不同生姜品种超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)、苯丙氨酸解氨酶(PAL)、几丁质酶(CHI)、β-1,3-葡聚糖苷酶(GLU)活性及次生抗菌物质含量、超氧阴离子(O₂-·)、过氧化氢(H₂O₂)、丙二醛(MDA)含量等的影响,结合抗病系数、主成分分析、聚类分析、相关分析和隶属函数对5个生姜品种幼苗的抗病性进行综合评价。【结果】山东大姜病情指数最低(4.0)、发病率最低(4%)、发病周期最长(24 d),表现出较强的抗病性;罗平小黄姜病情指数最高(45.2)、发病周期最短(7 d),表现抗病性较弱。不同生姜品种幼苗接菌后的生理生化指标均出现不同程度的变化,其中,山东大姜的抗氧化酶(SOD、POD、CAT)活性、防御酶(PAL、CHI、GLU)活性和次生抗菌物质(总酚、类黄酮、木质素)含量均高于其他品种,O₂-·、H₂O₂和MDA含量低于其他品种,而罗平小黄姜呈现相反趋势。主成分分析将12个单项生理生化指标转换为3个独立综合指标,聚类分析将5个生姜品种划分为3类,结合隶属函数综合评价结果,5个生姜品种抗病性强弱表现为:山东大姜>凤头姜>竹根姜>贵州黄姜>罗平小黄姜,与人工接种试验中5个品种的发病程度一致。相关分析结果显示,SOD和类黄酮与抗病性综合评价值(D值)呈极显著正相关(P<0.01)。【结论】5个生姜品种中山东大姜抗病性最强,罗平小黄姜抗病性最弱;SOD和类黄酮可作为生姜枯萎病抗性鉴定的主要参考指标。     

Improvement in winter wheat productivity through regulating PSII photochemistry,photosynthesis and chlorophyll fluorescence under deficit irrigation conditions

Deficit irrigation is critical to global food production, particularly in arid and semi-arid regions with low precipitation. Given water shortage has threatened agricultural sustainability under the dry-land farming system in China, there is an urgent need to develop effective water-saving technologies. We carried out a field study under two cultivation techniques: (1) the ridge and furrow cultivation model (R); and (2) the conventional flat farming model (F), and three simulated precipitation levels (1, 275 mm; 2, 200 mm; 3, 125 mm) with two deficit irrigation levels (150 and 75 mm). We demonstrated that under the ridge furrow (R) model, rainfall harvesting planting under 150 mm deficit irrigation combined with 200 mm simulated precipitation can considerably increase net photosynthesis rate (P_n), quantum yield of PSII (ΦPSII), electron transport rate (ETR), performance index of photosynthetic PSII (F_v/F_m′), and transformation energy potential of PSII (F_v/F_o). In addition, during the jointing, anthesis and grain-filling stages, the grain and biomass yield in the R model are 18.9 and 11.1% higher than those in the flat cultivation model, respectively, primarily due to improved soil water contents. The winter wheat fluorescence parameters were significantly positively associated with the photosynthesis, biomass and wheat production. The result suggests that the R cultivation model with irrigation of 150 mm and simulated precipitation of 200 mm is an effective planting method for enhancing P_n, biomass, wheat production, and chlorophyll fluorescence parameters in dry-land farming areas.     

Tomato mottle mosaic virus: Characterization,resistance gene effectiveness,and quintuplex RT-PCR detection system

Tomato mottle mosaic virus (ToMMV), an economically important species of the genus Tobamovirus, causes significant loss in yield and quality of tomato fruits. Here, we identified the Shandong isolate of ToMMV (ToMMV-SD) collected from symptomatic tomato fruits in Weifang, Shandong Province of China. ToMMV-SD caused symptoms such as severe mosaic, mottling, and necrosis of tomato leaves, yellow spot and necrotic lesions on tomato fruits. The obtained full genome of ToMMV-SD was 6 399 nucleotides (accession number MW373515) and had the highest identity of 99.5% with that of isolate SC13-051 from the United States of America at the genomic level. The infectious clone of ToMMV-SD was constructed and induced clear mosaic and necrotic symptoms onto Nicotiana benthamiana leaves. Several commercial tomato cultivars, harboring Tm-2² resistance gene, and pepper cultivars, containing L resistance gene, were susceptible to ToMMV-SD. Plants of Solanum melongena (eggplant) and Brassica pekinensis (napa cabbage) showed mottling symptoms, while N. tabacum cv. Zhongyan 100 displayed latent infection. ToMMV-SD did not infect plants of N. tabacum cv. Xanthi NN, Brassica rapa ssp. chinensis (bok choy), Raphanus sativus (radish), Vigna unguiculata cv. Yuanzhong 28-2 (cowpea), or Tm-2² transgenic N. benthamiana. A quintuplex RT-PCR system differentiated ToMMV from tomato mosaic virus, tomato brown rugose fruit virus, tobacco mosaic virus, and tomato spotted wilt virus, with the threshold amount of 0.02 pg. These results highlight the threat posed by ToMMV to tomato and pepper cultivation and offer an efficient detection system for the simultaneous detection of four tobamoviruses and tomato spotted wilt virus infecting tomato plants in the field.     

Neopestalotiopsis eucalypti,a causal agent of grapevine shoot rot in cutting nurseries in China

Grapevine (Vitis vinifera L.) is an economically important fruit crop in the world, and China ranks first in the production of grapes with approximately 15% of the world's total yield. However, diseases that cause the death of grapevine shoots pose a severe threat to the production of grapes. In this study, the fungus Neopestalotiopsis eucalypti was identified as a causal pathogen of grapevine shoot rot based on the morphology of conidia and a phylogenetic analysis. The phylogenetic analysis was performed with three isolates based on the combined sequence of internal transcribed spacer (ITS) region of ribosomal DNA, part of the translation elongation factor 1-alpha (Tef) and the β-tubulin (Tub2) genes. The three isolates were all identified as N. eucalypti. Pathogenicity tests of the three fungal isolates were conducted on grapevines shoots in vitro and in vivo. The results showed that all three fungal isolates caused severe rot lesions on the inoculated grapevine shoots, and N. eucalypti was re-isolated from the inoculated grapevine shoots. Therefore, N. eucalypti was confirmed as a causal agent of the grapevine shoot rot. This is the first report of N. eucalypti causing grapevine shoot disease in China.