Diaporthe spp. (including their asexual Phomopsis states) are important plant pathogens found widely in different host plants, such as pathogens, endophytes or saprobes. In this study, a new species of Diaporthe causing fruit rot in kiwifruit, Diaporthe xiuwenensis sp. nov was isolated from kiwifruit with apparent soft rot symptoms during storage in Guizhou Province, China. Identification was made using morphology and comparison of DNA sequence data of the nuclear ribosomal ITS (internal transcribed spacer) + tef1 (translation elongation factor-1 alpha) + tub2 (beta-tubulin) + cal (calmodulin) + his (histone H3) gene regions. This study further described a new Diaporthe species and discussed its morphology and phylogenetic relationships with other Diaporthe species. Koch's postulates tests identified the causative agents of kiwifruit soft rot. The results of pathogenicity test showed that Diaporthe xiuwenensis sp. nov was pathogenic to kiwifruit, and the Koch's postulates were completed.

Kiwifruit canker and brown spot are significant diseases affecting kiwis, caused by Pseudomonas syringae pathogenic variations (Pseudomonas syringae pv. Actinidiae (Psa)) and Corynesporapolytica (Corynespora cassiicola). At present, the research on canker disease and brown spot disease mainly focuses on the isolation and identification of pathogenic bacteria, drug control, resistance gene mining and functional verification. Practice has proved that breeding disease resistant varieties are an effective method to control canker disease and brown spot disease. However, most existing cultivars lack genes for canker and brown spot resistance. Wild kiwifruit resources in nature exhibit extensive genetic diversity due to prolonged natural selection, containing numerous resistance genes. But, due to insufficient understanding of the resistance of most kiwifruit varieties (lines) to canker disease and brown spot disease, some high-quality resources have not been fully utilized. The incidence of canker and brown spot of 18 kiwifruit cultivars (lines) was measured by inoculating isolated branches and leaves, and their resistance to canker and brown spot was analyzed according to the length, disease index, mean diameter, and systematic clustering. The results were as follows: Among 18 different kiwifruit varieties (lines) for canker disease, there were two highly resistant materials, eight disease-resistant materials, four disease-susceptible materials, and two highly susceptible materials. Moreover, regarding brown spot disease, there were one highly resistant material, five disease-resistant materials, four susceptible materials, and three highly susceptible materials. Furthermore, four resources were resistant to both diseases. The outcomes provided a theoretical basis for breeding kiwifruit against canker and brown spot.

为了明确氯吡脲(CPPU)和糖醇螯合钙对猕猴桃果实产量和品质的影响,筛选出改善猕猴桃果实品质的最优处理,以红阳猕猴桃为试验材料,于果实膨大期,采用不同浓度CPPU与糖醇螯合钙及两者组合浸果,果实成熟期测定果实品质各项指标,并对指标进行主成分分析.结果表明:CPPU与糖醇螯合钙处理会不同程度地增加果心空腔率与畸形果率,降低裂果率;不同浓度CPPU与糖醇螯合钙及两者组合对猕猴桃果实品质及产量的影响不同,C4处理(10.0 mg/L CPPU+1 000 mg/L糖醇螯合钙)的增产效果最显著,其次是A3处理(7.5 mg/L CPPU),增产率分别达96.52％与84.88％;基于主成分分析的综合评价得出,A3处理(7.5 mg/L CPPU)对果实产量的提升及品质的改善效果最优,其次是C4处理,其余处理综合得分均高于CK(清水).综上,氯吡脲和糖醇螯合钙处理能在保证猕猴桃果实品质的前提下,显著提高果实产量.

【Objective】To understand the effects of different disease resistance inducers on the field control effect, leaf nutrient content, fruit yield and quality of kiwi brown spot, in order to screen out the best disease resistance inducers for the prevention and control of kiwi brown spot, and provide technical support for the green prevention and control of kiwi brown spot.【Method】Taking ‘Hongyang’ kiwifruit as the research object, field experiments were carried out with single application and mixed application of different disease resistance inducers, and the incidence, disease index,control effect and leaf nutrient content of brown spot disease in the field were determined after the investigation, and the relevant indicators of the fruit were measured and comprehensively evaluated at the ripening stage.【Result】There were significant differences in the effect of different disease resistance inducers on the prevention and treatment of brown spot. Treatment 6（Artaine + Heyi Sea Star） had the best prevention and control effect, and its prevention and treatment effects were 52.72%, 78.39% and 73.28% in 10, 20 and 30 days after the medication, followed by the prevention and treatment effect of treatment 5（aminooligosaccharin + Heyi Sea Star）. Compared with CK（methyltobutazine）, the contents of nitrogen（N） and potassium（K） in leaves were increased by each treatment of disease resistance inducer in 10 days after treatment. The N content of leaves changed greatly in 10, 20 and 30 days after the medication, and the overall content showed a significant downward trend. Among them, the leaf N content of treatment 6（Artaeling + Heyi Sea Star） was the highest. The overall variation of K content in leaves in 10, 20 and 30 days after medication treatment was small. In 10 days after the medication, the K content of the leaves of treatment 5（aminooligosaccharin + Heyi Sea Star） was the highest. At 20 and 30 days after the medication, the K content of the leaves of treatment 6（Artaelin + Heyi Sea Star） was the highest. However, the inducer failed to effectively increase the phosphorus（P） content of leaves. The correlation analysis results showed that the N content of leaf was significantly negatively correlated with the incidence and disease index of brown spot, indicating that the disease resistance inducer mainly enhanced its disease resistance ability by inducing the absorption of leaf N pigment. The comprehensive evaluation results showed that treatment 5（aminooligosaccharin + Heyi Sea Star） had the best effect on fruit yield and quality, followed by treatment 6, CK had the worst improvement effect.【Conclusion】Treatment 5（aminooligosaccharin + Heyi Sea Star） had significant effects on immune induction, fruit yield and fruit quality improvement, and aminooligosaccharin + Heyi Sea Star could be used as the best disease resistance inducer in green control of kiwifruit brown spot.

通过对146份贵州猕猴桃种质资源表型遗传多样性的研究,为贵州猕猴桃品种选育打下基础,对146份不同区域猕猴桃种质资源48个质量性状进行描述、赋值,并结合单果重等9个数量性状,使用主成分分析、聚类分析等进行综合评价.结果表明,57个质量性状和数量性状均存在丰富的多样性和变异性;146份资源的Shannon's遗传多样性指数为1.28,从各区域遗传多样性指数可以看出,演化进程上从遗传多样性指数高的地区向低的地区进行演化;主成分分析结果显示,前14个主成分累计贡献率达75.076％,其中前6个主成分的累计贡献率达51.911％,为提高品种选育和评价的工作效率,可将猕猴桃种质资源的描述指标简化为新梢被毛、果实被毛等18个性状;聚类结果显示,贵州猕猴桃种质的欧氏距离为1.00～25.00之间,在D=19.25处,146份野生猕猴桃分为6个类群.本文有效揭示了贵州省猕猴桃种质资源丰富的多样性和亲缘关系,明确野生猕猴桃各种群间性状区分特征和部分优良资源性状的地理分布,对贵州猕猴桃种质资源的种质鉴定、保护利用和品种选育有重要意义.

为了明确红阳猕猴桃褐斑病主要病原菌及适宜的防治药剂,采集贵州省六盘水市水城区红阳猕猴桃褐斑病的标样,通过组织分离、纯化,致病性检测,形态学观察,分子检测及系统发育分析确定病原菌种类,并采用菌丝生长速率法测定6种药剂对该病原菌生长的抑菌效果.结果显示:分离纯化得到的病原菌的形态特征与多主棒孢菌Corynespora cassiicola形态特征相似,将分离菌株rDNA-ITS序列与GenBank中相关菌株的ITS序列进行同源性比对,发现该菌株与黄瓜多主棒孢霉菌的相似性达到96％以上.室内药剂筛选结果表明:50％多菌灵可湿性粉剂对病原菌的抑制效果最好,最低浓度可抑制菌丝生长;其次为40％戊唑醇悬浮剂、80％乙蒜素乳油、30％吡唑嘧菌酯乳油和80％代森锰锌可湿性粉剂,抑菌效果较好,EC50值分别为0.000 2、0.074 3、0.087 1、和0.502 5 mg/L.红阳猕猴桃褐斑病主要由多主棒孢菌Corynespora cassiicola侵染引起,以上5种药剂均可作为防治红阳猕猴桃褐斑病的有效药剂.

[目的]探究猕猴桃苗不同生长时期根际土壤细菌群落组成及土壤养分间的相互关系.[方法]采用Illumina-MiSeq高通量测序研究猕猴桃苗不同生长阶段(生长前期、速生期、生长后期)的根际微生物群落结构组成及其多样性.运用CANOCO和R语言软件对土壤细菌群落与土壤环境因子间关系进行RDA和相关性热图分析.[结果]随着猕猴桃苗不断生长,土壤pH、速效N、速效P均出现先升高后降低趋势,速效K则随着猕猴桃苗生长逐渐降低;高通量测序共得到25 822个细菌OTUs,分属于28门53 纲 172 科 226 属,其中变形菌门(Proteobacteria,35.84％)、酸杆菌门(Acidobacteria,17.69％)、放线菌门(Actinobacteria,12.6％)是猕猴桃苗不同生长时期土壤的优势菌门;土壤细菌群落多样性指数和丰富度指数在猕猴桃生长初期与速生期和生长后期有显著差异;不同生长时期相对丰富度大于1％的菌门中,Cyanobacteria随着猕猴桃苗不断生长而逐渐减少,Actinobacteria、Latescibacte-ria、Acidobacteria的相对含量先升高后降低.[结论]猕猴桃苗根际土壤细菌群落组成和根际土壤养分发生变化受猕猴桃苗不同生长时期的影响,土壤根际主要养分和pH是影响土壤根际细菌群落结构比例变化的重要因子.

为探明不同有机肥料对"红阳"猕猴桃叶片营养及果实品质的影响,参照贵州省水城县中等猕猴桃产量及肥力水平,设置对照、复合肥以及4种有机肥(牛粪,羊粪,油枯有机肥,微生物菌肥);共6个处理,分别于"红阳"猕猴桃萌芽期、开花期、第1次果实膨大期、第2次果实膨大期及成熟期采集样品进行测试分析.结果表明:"红阳"猕猴桃叶片干物质N和K含量表现为微生物菌肥>油枯有机肥>复合肥>羊粪>牛粪>对照;P含量表现为微生物菌肥>油枯有机肥>羊粪>复合肥>牛粪>对照;果实N、P、K以及水溶性总糖、干物质、可溶性固形物含量表现为微生物菌肥>油枯有机肥>羊粪>牛粪>复合肥>对照;维生素C含量表现为牛粪>油枯有机肥>微生物菌肥>羊粪>复合肥>对照;对于果实总酸度来说,对照以及复合肥处理显著高于有机肥处理,但牛粪、羊粪、有机肥以及微生物菌肥之间的差异不明显.在"红阳"猕猴桃整个生育期,叶片N含量缓慢降低,其中复合肥处理的降幅最大,而所有处理叶片P和K含量的降幅都较大,采摘期比萌芽期分别降低80％和70％以上.

为探究猕猴桃苗不同生长期与根际土壤真菌群落结构、多样性及土壤养分之间的相互关系,利用Illumina MiSeq高通量测序研究猕猴桃苗3个不同生长阶段(生长前期、速生期、生长后期)根际微生物群落结构组成及其多样性,运用CANOCO和R语言软件对土壤真菌群落与土壤环境因子间关系进行RDA和相关性分析.结果表明:猕猴桃苗不同生长期土壤性质均存在显著差异,随着猕猴桃苗根系不断生长,土壤pH、速效氮、有效磷出现先升高后降低趋势,速效钾则出现先降低后升高;高通量测序共得到1 776个真菌OTUs,分属于10门22纲151科157属;方差分析表明各生长时期多样性指数(Shannon)差异显著;序列分析发现,子囊菌门(Ascomycota,丰度72.69％)、被孢霉门(Mortierellomycota,18.80％)、罗兹菌门(Rozellomycota,6.68％)、担子菌门(Basidiomycota,4.28％)和球囊菌门(Glomeromycota,1.45％)是猕猴桃苗不同生长期的优势菌门(丰度大于1％的视为优势菌门);对不同生长时期优势菌门所占比例进行分析,发现子囊菌门出现先升高后降低,被孢霉门、罗兹菌门和担子菌门则出现先降低后升高,球囊菌门相对丰度随着猕猴桃根系生长逐渐升高;RDA分析结果显示,有效磷和pH与子囊菌门显著正相关,速效钾与子囊菌门显著负相关,有机碳与壶菌门(Chytridiomycota)显著负相关.综上,猕猴桃苗在不同生长时期根际土壤真菌群落结构不同,速生期根际真菌种群最丰富,生长后期最少;子囊菌门、被孢霉门、罗兹菌门和担子菌门始终是猕猴桃苗生长期根际优势菌群;根际真菌群落组成和土壤养分变化受猕猴桃苗不同生长时期的影响,其中土壤有效磷和速效钾是影响子囊菌门群落组成的重要因子.

This study aimed to investigate the response of sediment microbial communities (including bacteria and archaeal groups) in Caohai Lake to anthropogenic activities. The sediment samples were collected from the regions with high anthropogenic interference and low anthropogenic interference. Their physicochemical properties and enzyme activities were analyzed, and the bacterial and archaeal communities were investigated using high-throughput sequencing technology. The results showed that the physicochemical characters changed by anthropogenic activities were the important factors that influenced enzyme activities, alpha diversity, key functional taxa, and community structure. And the impact of anthropogenic activities on microbial communities might follow a non-linear pattern. Furthermore, few significant differences of alpha indices between the high and low disturbed areas, but clear differences of microbial community composition analysis and beta-diversity analysis were observed. The hypothesis was proved that the intensity of anthropogenic impacts in Caohai had not reached the potential thresholds. The best distinguish biomarkers between the two areas and the most related key nodes among the network did not always have a high microbial abundance. The anthropogenic activities might influence the microbial community by affecting a small number of the key taxon in the ecological network. These findings provided a valuable understanding of how sediment microorganisms respond to anthropogenic activities in Caohai Lake.

为了筛选百香果最佳施肥模式,本研究设置5种不同施肥处理:不施肥(CK)、氮磷钾肥(NPK)、有机肥(M)、氮磷钾与有机肥配施(MNPK)、高量氮磷钾与高量有机肥配施(2MNPK),通过分析比较百香果的生长状况、产量、营养成分差异,进一步采用多元统计分析方法进行主成分分析及综合评价.结果发现:与CK处理相比,NPK、MNPK、2MNPK处理百香果生长最快,尤其在定植66 d后效果最明显.NPK、M、MNPK和2MNPK处理均提高了百香果产量及品质,MNPK、2MNPK处理效果最明显.与CK处理相比,NPK、M、MNPK和2MNPK处理百香果产量分别增产112.86％、74.40％、191.00％、244.30％,维生素、矿质元素、糖类物质、氨基酸含量均有上升趋势,总酸呈下降趋势,2MNPK处理总氨基酸、必需氨基酸、风味氨基酸含量分别增加了82.40％、76.11％、76.89％,总酸含量降低了22.61％.对36种指标进行多元统计分析,从中提取了3个主成分因子,其累计方差贡献率为97.925％.建立综合评价模型计算综合总得分排序为2MNPK(123.04)>MNPK(39.06)>M(-12.01)>NPK(-45.36)>CK(-104.73).说明在这5种施肥处理中,2MNPK处理是百香果种植过程中最佳施肥模式.该研究可以为百香果大面积推广应用奠定理论基础.

为明确贵州省猕猴桃主栽品种果实腐烂病病原菌的种类,并为田间果实病害防治提供理论依据,以2018—2019年在贵州省水城县和修文县两大主产区采集的红阳和贵长猕猴桃病果为试验材料,通过病原菌的分离、形态学观察、分子检测和致病性检测,研究病果中病原真菌的种类,并对其序列进行系统发育分析.结果表明,分离到4类病原菌:拟茎点霉菌(Phomopsis cauloides)、镰刀菌(Fusarium)、交链格孢菌(Alternaria alternata)、黄瓜织球壳菌(Plectosphaerella cucumerina),其检出率分别为10％、60％、20％、10％.其中,黄瓜织球壳菌侵染引起猕猴桃果实病害在国内尚属首次报道.

[目的]筛选能促进猕猴桃生长和提高果实品质的最佳处理,为猕猴桃合理施肥提供参考.[方法]以'红阳'Actinidia chinensis'Hongyang'猕猴桃为试材,于壮果期,采用沟施方式,在施用复合肥的基础上,增施有机肥和菌肥,测定和统计各处理下猕猴桃果实的外观性状、内在品质及果实采后软腐病发生率.[结果]与对照相比,增施有机肥和菌肥后,猕猴桃果实的横纵径、果形指数无显著差异,各处理的单果质量不同程度有所增加,但与对照均无显著差异.其中复合肥、海藻甲壳素混施处理(处理2)中猕猴桃单果质量增加幅度最大,较对照增加了23.82％.增施有机肥和菌肥处理对果实中微量元素含量的影响显著.增施有机肥和菌肥后,除硼含量外,果实中其他元素含量均显著高于对照.其中,复合肥、菌肥、黄腐酸钾、海藻甲壳素混施处理(处理6)中果实钙和锌含量高于其他处理及对照,分别比对照显著增加了5.07％与5.44％;果实中锰含量最高的是复合肥、菌肥、黄腐酸钾混施处理(处理4),镁含量最高的是处理2,分别比对照显著增加了62.16％与72.13％.增施有机肥和菌肥后,果实的可溶性固形物含量、可溶性糖含量、可溶性蛋白含量和糖酸比均高于对照,维生素C含量除了处理2,其余处理均高于对照,处理6的果实维生素C含量最高,菌肥、黄腐酸钾、海藻甲壳素混施处理(处理5)次之,分别较对照高出26.86％和24.21％.增施有机肥和菌肥后,果实软腐病发病率均比对照低,且差异显著.处理6的发病率最低,较对照低41.07％.[结论]在施用复合肥的基础上,增施有机肥和菌肥可较好地改善猕猴桃果实的外观性状和内在品质,并减少果实病害发生,以复合肥、菌肥、黄腐酸钾、海藻甲壳素混施的增质效果最好.

目的 探索不同自发气调袋在低温下对百香果的贮藏效果.方法 将"台农1号"百香果作为实验材料,使用不同自发气调袋对其进行包装处理,贮藏于(8±0.3)℃的环境内30 d,每5 d进行1次生理和营养指标的测定,研究贮藏期间各自发气调袋内百香果的品质变化,以期探寻"台农1号"百香果的最佳贮藏包装条件.结果 随着贮藏时间的延长,PE袋内的CO2浓度逐渐升高,O2浓度逐渐下降,相较于微孔袋,PE袋推迟了贮藏期间果实的呼吸高峰,延缓了可溶性固形物、维生素C、总黄酮和多酚含量的下降,维持了较高的果皮硬度;而不同厚度的PE袋之间,果实品质也不同,贮藏到30 d时,PE30果皮硬度(16.17 g)、可溶性固定物(15.29％)、维生素C(1.71 mg/kg)、总黄酮(2.14 mg/kg)含量均最高.结论 "台农1号"百香果在低温下采用PE30自发气调袋进行包装,其贮藏效果最佳.

猕猴桃褐斑病是猕猴桃叶部主要病害,也可以发生于果实上,严重时可以引起果园叶片大面积提早脱落,大量秋稍萌发,对猕猴桃的产量和品质造成一定影响.根据近几年国内外报道,对猕猴桃褐斑病的发病症状、病原及寄主,发生规律及防治进行简要综述,并展望了猕猴桃褐斑病的研究方向.

为了探讨不同砂土类型和不同埋藏方法对猕猴桃种子萌发及幼苗生长的影响,以梵净山野生猕猴桃种子为供试材料,研究了不同处理对猕猴桃种子萌发率、出苗率以及幼苗生长的影响.结果 表明:石英砂与腐殖土混合的砂土最适合猕猴桃种子萌发,平均萌发率达到84％,室内埋藏后的种子更有利于种子出苗,出苗率达到81.6％.方差分析结果表明不同砂土类型和不同埋藏方法对种子萌发率和出苗率均显著影响,该研究结果为猕猴桃种子资源保存和种苗繁育提供了理论依据.

本实验室前期在高渗条件下筛选得到8个转录因子,其中ACZ (SI65-00458)基因表达量最高,推测ACZ基因在冠突曲霉中参与响应渗透压及调控孢子的产生.因而本研究利用同源重组原理,构建了ACZ基因敲除载体,通过农杆菌介导转化,筛选获得了ACZ敲除菌株,将敲除株分生孢子液接种在含不同浓度NaCl的MYA培养基上,以野生型冠突曲霉为对照,观察敲除株与野生型菌株的区别.研究发现:培养基中不加NaCl时色素有明显变化,野生型为黄色,敲除株为褐黄色;在低渗透压下,菌落边缘不规则;在高渗透压下,敲除株产生的分生孢子数量为野生型的4倍多;无论是在27℃还是在37℃培养,敲除株在低渗及高渗条件下,菌落直径都较野生型小,菌丝较稀疏.表明敲除ACZ基因会影响冠突曲霉菌丝生长、色素的合成及孢子的产生.

2015-2017年作者采用随机普查和定点调查方式调查贵州省蓝莓Vaccinium sp.主要栽培区病害种类.结果发现:贵州蓝莓叶、茎、花共有7种真菌病害,其中蓝莓褐斑病(Pseudocercospora sp.)、蓝莓炭疽病(Colletotrichum sp.)、蓝莓茎枯病(Ophiobolus sp.)、蓝莓枯萎病(Phomopsis sp.)值得后期密切关注.

The metabolites of mycelial samples of ΔveA mutant and wild-type strain E4 of Aspergillus cristatus with 48 hour incubation period were identified by gas chromatography-mass spectrometry (GC-MS).A total of 99 metabolites were identified,including 41 significantly differential metabolites.Among them,20 and 21 metabolites were much increased in the wild-type E4 and ΔveA,respectively.Statistical analysis revealed there were 623 individual metabolite having correlation,of which 313 were positive and 310 negative.These metabolites include organic acids,amino acids,carbohydrates,alcohols and oxylipin,while organic acids and amino acids predominate.This result lays the foundation for further study on the correlation between metabolite and sporulation in A.cristotus.

veA belongs to the velvet regulatory system that regulates the development and secondary metabolism of many fungi. To identify the function of veA in Aspergillus cristatus, veA deletion mutants were constructed by homologous recombination via Agrobacterium tumefaciens-mediated transformation. Deletion of veA led to increased conidial production and reduced sexual sporulation. The regulatory role of veA in A. cristatus was not light-dependent, and this differed from its role in other Aspergilli. Furthermore, veA deletion mutants were more sensitive to environmental stressors, including salt, osmotic pressure, temperature and pH. In contrast, deletion of veA resulted in increased resistance to oxidative stress. veA also affected aerial vegetative growth. Transcriptomic analysis of the veA-null mutant and wild type indicated that most asexual and sexual development genes were upregulated and downregulated, respectively. These findings confirmed that veA has a positive effect on sexual development but represses conidial formation. Overall, these results suggested that the veA gene plays a critical role in maintaining a developmental balance between asexual and sexual sporulation and is involved in vegetative growth and environmental stress response in A. cristatus.