嫁接对茄子连作土壤生物活性及种子萌发和幼苗生长的影响

研究了嫁接茄根系分泌物对黄萎菌菌丝生长的影响,嫁接对茄子根际土壤黄萎菌数量、酶活性和微生物种群数量影响及嫁接正重茬土壤水提液对茄子种子萌发和幼苗生长的影响。结果表明：在培养4～5 d,嫁接茄根系分泌物显著抑制了黄萎菌菌落生长;嫁接茄土壤黄萎菌数量在不同时期均显著低于对照;总体上,嫁接茄根际脱氢酶、过氧化物酶和多酚氧化酶较对照有所增强;嫁接还表现出根际微生物总量、细菌、放线菌数量增加,真菌数量减少,细菌数比真菌数和放线菌数比真菌数增加;嫁接茄正重茬土壤水提液表现出促进茄子发芽和幼苗生长(根长除外),重茬自根茄土壤水提液抑制了茄子发芽和幼苗生长。     

嫁接茄根系分泌物与抗黄萎病的关系及其组分分析

探讨了嫁接对茄子土传病害黄萎病抗性的影响,采用浸根法收集茄子根系分泌物,并利用气-质联用技术检测了根系分泌物的组分和含量,测定了茄子根系分泌物及其模拟化感物质对黄萎菌菌丝生长的化感作用,分析嫁接后茄子根系分泌物的组成差异,探讨了嫁接茄抗病性与其根系分泌物成分的关系。结果表明:嫁接使茄子发病率显著降低。土壤黄萎菌数量随发育时期的推进而显著低于自根茄。嫁接茄根系分泌物对黄萎菌菌丝生长有一定的抑制作用。GC-MS检测结果显示,嫁接换根使茄子根系分泌物的组成成分发生一定的改变,嫁接根系分泌物中检测到了烃类、酯类、醇类、酚类、酮类、苯类和胺类物质,其中苯、醇和胺类在ZG中未检测到。但嫁接茄也缺少自根茄中检测到的炔类和菲啶。酚类公认的化感物质含量表现出自根茄大于嫁接茄。模拟物质对黄萎菌菌丝生长的影响表明:邻苯二甲酸二丁酯表现出在0.05 0.5 mmol.L-1浓度范围随浓度增加抑制作用增强,后随浓度增加抑制作用有所减弱。而豆蔻酸和棕榈酸均表现出随浓度增加先促进后抑制黄萎菌菌丝的生长。     

嫁接茄子根系分泌物变化及其对黄萎菌的影响

采用番茄为砧木嫁接茄子,经过GC-MS检测,研究了黄萎菌胁迫前后嫁接对茄子植株根系次生代谢的影响。通过比较黄萎菌胁迫前后茄子根系次生代谢物质的变化,探讨了嫁接在胁迫前后对植株根系次生代谢的调节作用,并对嫁接茄根系分泌物中丁二酸二甲酯对茄子黄萎菌及茄子种子萌发、幼苗生长的化感效应进行了研究。结果表明,黄萎菌胁迫前,嫁接影响了根系的次生代谢物质的分泌,表现为物质种类和数量的增加,各类物质相对含量改变。嫁接茄子根系分泌物中检测出9大类、66种物质,比自根茄处理多出4大类、33种物质。黄萎菌胁迫时,嫁接茄子田间表现出明显的抗病性;进一步对根系分泌物进行检测发现,嫁接茄根系分泌物中物质种类和相对含量与自根茄处理相比均有显著差别;嫁接茄根系分泌物中烃和酚醇类物质相对含量分别增加了3.25%和0.07%,苯类、茚类和脂肪酸酯类物质相对含量降低,降幅分别为2.62%、0.26%和0.07%。新出现了胺类物质,芴类物质。黄萎菌胁迫前后茄子根系次生代谢物质成分同样发生了变化。与接菌前嫁接茄子植株根系分泌物相比,接菌后嫁接处理的苯类、茚类、酚醇类和胺类物质相对含量增加,增幅分别为22.07%、1.72%、1.21%和0.34%;烃类和脂肪酸酯类物质相对含量降低了1.28%和21.75%;酮类、咔唑类和芴类物质消失。新增物质中以丁二酸二甲酯的相对含量最高,达14.38%。随后的生物检测结果显示,丁二酸二甲酯能够提高茄子田间抗病性,对黄萎菌菌丝生长起化感抑制作用,并促进了茄子种子的萌发和幼苗的生长,随着处理浓度升高作用效果增强,并在1 mmol/L处理时达到最佳作用效果。     

不同抗病性茄子根系分泌物对黄萎菌的化感作用

以不同抗病性茄子为试材,即抗病类型Solanum tor、S.sis,耐病类型立原紫茄,感病类型西安绿茄,研究了不同茄子品种抗黄萎病特性,根际微生物结构与黄萎菌数量的变化,茄子根系分泌物对黄萎菌的化感作用,并利用GC-MS对根系分泌物的成分进行了鉴定。结果表明：抗病类型的根系分泌物既可以直接影响黄萎菌的生长、发育,又可以通过调节土壤微生物种群结构间接影响黄萎菌的生长,达到抗病效果。而感病类型则正相反。推断这可能是因为抗病类型根系分泌物中存在醇类、胺类、吡喃类、芴类等特异物质,而感病类型根系分泌物中酮类、酚类、酯类、酚酸类物质种类或含量较高。     

不同连作年限土壤对茄子土壤生物学活性的影响及其嫁接调节

探讨了不同连作年限土壤对茄子根际土壤微生物数量、土壤酶活性的影响及其相关性和嫁接对重要相关因子的调节作用。结果表明:随着连作年限的增加,茄子根际土壤细菌数量和放线菌数量呈下降趋势,真菌数量呈增加趋势;根际土壤中的过氧化氢酶、过氧化物酶、多酚氧化酶、蛋白酶的活性呈降低趋势;土壤微生物数量和土壤酶活性之间有显著的相关性。其中过氧化氢酶、过氧化物酶、多酚氧化酶、蛋白酶是影响根际土壤中细菌和放线菌数量的主要因子,蛋白酶是影响根际土壤中真菌数量的主要因子。嫁接能提高土壤酶活性,缓解连作土壤带来的胁迫。     

刺萼龙葵不同生育期根际土壤酶活性和真菌多样性变化

分析入侵植物刺萼龙葵(Solanum rostratum)在不同生育期根际土壤真菌多样性及土壤酶活性变化,探讨刺萼龙葵入侵对土壤微生态的影响机制,以期寻找防治刺萼龙葵的最佳时期。本试验利用土壤平板法对刺萼龙葵根际土壤真菌进行培养,共鉴定真菌22个属,并且从生长初期到开花期土壤真菌多样性显著增加;从生长初期到开花期根际土壤过氧化氢酶、过氧化物酶、多酚氧化酶、脲酶、脱氢酶、转化酶和蛋白酶活性变化显著;Pearson相关分析发现,刺萼龙葵从生长初期到开花期根际土壤真菌多样性与过氧化物酶活性相关性并不显著,而与其余6种土壤酶活性具有显著的相关性;主成分分析发现,刺萼龙葵开花期、现蕾期和四叶期与空白土壤和生长初期发生了显著的变化。因此,随着刺萼龙葵的生长,其根际土壤真菌多样性及土壤酶活性均发生改变。     

嫁接对铜胁迫下黄瓜根际土壤微生物特性和酶活性的影响

采用盆栽试验方法,研究了嫁接（以黑籽南瓜为砧木）对铜胁迫下黄瓜根际土壤微生物生物量、微生物种群数量和土壤酶活性的影响.结果表明： 铜胁迫下黄瓜根际土壤微生物生物量碳（MBC）和微生物生物量氮（MBN）含量显著下降,基础呼吸和代谢熵显著上升,但嫁接黄瓜根际土壤MBC和MBN含量显著高于自根黄瓜,而基础呼吸和代谢熵则显著低于自根黄瓜.铜胁迫下,根际土壤放线菌和自生固氮菌的数量显著下降,真菌数量显著上升,而细菌数量变化不显著;嫁接黄瓜根系土壤细菌、放线菌、自生固氮菌的数量显著高于自根黄瓜,而真菌数量显著低于自根黄瓜.嫁接黄瓜根际土壤脲酶、磷酸酶、蔗糖酶、过氧化氢酶活性在铜胁迫下显著高于自根黄瓜.试验结果证明嫁接使铜胁迫下黄瓜根际土壤微生物环境和酶活性得到了改善和提高,从而提高了黄瓜植株对铜胁迫的抵抗力.     

伊贝根际微生物分布与西贝素的相关性

分析了伊犁贝母不同生育期(返青期、显蕾期、盛花期和收获期)根际、非根际可培养土壤微生物动态变化,探讨了根际土壤微生物与西贝素含量之间的相关性。结果表明,在伊犁贝母的生长发育期,根际和非根际土壤微生物数量的分布规律是:细菌放线菌真菌。除了显蕾期非根际真菌数量大于根际以外,其他生长时期根际细菌、放线菌和真菌数量远远大于非根际,细菌根际效益明显。盛花期根际土壤中细菌和真菌数量达到了最多。土壤真菌与西贝素含量呈极显著正相关。     

AM真菌对青枯菌和根际细菌群落结构的影响

利用传统的平板培养与DGGE相结合的技术手段,研究了接种AM真菌对番茄根际土壤中的青枯菌和细菌群落结构的影响。结果表明,菌根根际土壤中的细菌总量和总DNA量都高于非菌根根际土壤,其中前者的青枯菌种群数量比后者低60倍;DGGE图谱也证实了AM真菌对青枯菌的抑制效应,还揭示出接种AM真菌对根际土壤中细菌群落结构所产生的复杂的影响。文章对AM真菌抑制青枯菌的机制进行了探讨。     

土壤添加蛇床子(Cnidium monnieri)和苦参(Sophora flavescens)对茄子黄萎病及根际微生物的化感影响

研究了蛇床子、苦参对茄子黄萎病菌的化感抑制作用及对茄子根际微生物数量的影响。结果表明,蛇床子、苦参的提取物抑制了茄子黄萎病菌菌丝生长,并随提取物浓度的增加抑制作用增强。在土壤中施入蛇床子、苦参提取物处理茄子苗后表现出一定的抗病效果,其中蛇床子处理的植株发病率最低。在土壤中添加蛇床子和苦参粉末,经腐解后,苦参处理的茄子植株株高、茎粗均高于对照处理,且二者处理后植株干物质含量高于对照。处理后茄子根际微生物中放线菌数量增加,真菌的数量变化差异不明显,细菌的数量相对减少,在初花期微生物的总量均高于初果期和定植期。     

Effect of Paenibacillus alvei, strain K165, on the germination of Verticillium dahliae microsclerotia in planta

Verticillium wilt is a devastating disease of a wide range of herbaceous and woody plant hosts, incited by the soilborne fungus Verticillium dahliae. In the present study, the effect of the potential biocontrol isolate Paenibacillus alvei, strain K165, on the germination of V. dahliae microsclerotia (msc) was investigated. Strain K165 was isolated from tomato root tips and its activity against V. dahliae has been shown in glasshouse and field experiments. In the present study, the application of K165 resulted in the reduction of msc germination of V. dahliae, in the root tips and the zone of elongation, of eggplants by 50% compared to the control treatment; whereas 10 and 12 cm away from root tips and in soil without plants the percentage of msc germination was reduced by 26% and 40%, respectively. However, K165 did not significantly affect the number and length of hyphae per germinated msc. In a split-root system, K165 triggered induced systemic resistance in eggplants against V. dahliae by reducing disease severity and msc germination by 27% and 20%, respectively. In addition, K165 colonised the rhizosphere of eggplants and soil in a population density of 5 and 3 log₁₀ cfu g⁻¹, 7 dpi, respectively. This is the first report of evaluating the direct/indirect effect of a rhizospheric bacterium on msc germination in the rhizosphere of eggplants, indicating that strain K165 reduces msc germination.     

Seedling vaccination by stem injecting a conidial suspension of F2, a non-pathogenic Fusarium oxysporum strain, suppresses Verticillium wilt of eggplant

Several bacterial and fungal strains have been evaluated as biocontrol agents (BCAs) against Verticillium dahliae. In these studies, the BCAs were applied as a root drenching inoculum; however, this application method may have an adverse effect on the native, beneficial for the plants, microbial community. In the present study, it was evaluated whether endophytic application by stem injecting a conidial suspension of the non pathogenic Fusarium oxysporum F2 strain, isolated from a V. dahliae suppressive compost amendment, could reduce significantly Verticillium wilt symptom development in eggplants. It was revealed that stem injection of F2 seven days before transplanting the seedlings to soil infested by V. dahliae microsclerotia resulted in reduced disease severity compared to the control treatment. To visualise F2 ramification into the plant vascular system eggplant stems were injected with an EGFP transformed F2 strain. It was shown that F2 colonises effectively the plant vascular tissues over a long period of time as it was assessed by F2 DNA levels. In parallel, qPCR analysis showed that the application of F2 reduced significantly the amount of V. dahliae DNA in the stem tissues compared to the control treatment.     

Assessing the Risk of Biological Control Agents on the Indigenous Microbial Communities: <Emphasis Type="Italic">Serratia plymuthica</Emphasis> HRO-C48 and <Emphasis Type="Italic">Streptomyces</Emphasis> sp. HRO-71 as Model Bacteria

The phytopathogenic fungus Verticillium dahliae Kleb. causes high yield losses in strawberry production. As effective chemical control of this fungus is no longer available, biological control based on natural antagonists might provide new control strategies. The aim of this study was to assess the impact of the two biological control agents S. plymuthica HRO-C48 and Streptomyces sp. HRO-71 on the rhizosphere community of the Verticillium host plant strawberry in field trials at two different sites in Germany. Therefore, we determined the abundances of culturable bacteria and investigated the community structure of the total rhizosphere microbiota by PCR-single strand conformation polymorphism analysis of the 16S rRNA and fungal ITS1 region. The abundances of culturable rhizobacteria on R2A medium as well as the proportion of in vitro Verticillium antagonists did not differ significantly. Additionally, no treatment specific differences were obtained in the composition of species of the non-target antagonistic bacteria in the rhizospheres. The culture-independent analysis revealed only transient differences between the bacterial communities not due to the treatments rather than to the plant growth stage. Fungal and bacterial community fingerprints showed the development of a microbiota, specific for a field site. However, no sustainable impact of the bacterial treatments on the indigenous microbial communities was found using culture-dependent and -independent methods.     

Quantitative assessment of the interaction between the antagonistic fungus Talaromyces flavus and the wilt pathogen Verticillium dahliae on eggplant roots

Quantitative aspects of the interaction between the antagonist Talaromyces flavus, the pathogen Verticillium dahliae and eggplant roots, were studied. When eggplant roots were inoculated with T. flavus, prior to the infection with the pathogen, the population density of T. flavus on V. dahliae-infected roots was at least 3 times higher than on healthy uninfected roots, and the proliferation of T. flavus on diseased eggplant roots was related to the severity of wilt symptoms, in the two levels of application of T. flavus studied. However, in all classes of disease severity tested (disease index, 0–3), the population density of T. Flavus on eggplant roots treated with 10⁶ ascospores g^–1 rooting mixture was significantly (p=0.05) higher than with 10⁵ ascospores g^–1. In roots treated with 10⁵ and 10⁶ T. flavus ascospores g^–1 rooting mixture, the population density of V. dahliae was reduced by 51% and 69%, respectively. When testing the relationships between the population density of V. dahliae in the roots and disease severity, no significant (p=0.05) difference was found between disease indexes 2 and 3. However, the density of V. dahliae on roots of plants with disease index 1 was significantly (p=0.05) lower than disease indexes 2 and 3. The positive relationship between the inoculum concentration of V. dahliae and the population density of T. flavus developed on eggplant roots was significant (p=0.001), linear, and highly correlated (r=0.945) on a logarithmic scale. In addition, the analysis of these data revealed a significant (p=0.05), high, negative and linear correlation (r=–0.985) between the log concentration of V. dahliae inoculum and the disease reduction achieved by T. flavus.     

根系土壤假丝酵母菌Candida sp. YIN9对大丽轮枝菌和全齿复活线虫的抑制作用

[目的] 研究黄萎病抗性棉(海 7124)根际土壤中酵母菌株对棉花黄萎病病原真菌大丽轮枝菌和全齿复活线虫的拮抗效果,为生物防治棉花黄萎病和全齿复活线虫提供理论依据。[方法] 通过镜检、糖发酵实验、碳源同化实验、26S rRNA测序对菌株的形态、生理生化特征及其系统发育关系进行鉴定,并利用七叶苷筛选、刚果红染色、平皿对峙实验、盆栽实验、平板生测实验测试其产酶活性以及抑制大丽轮枝菌和杀线虫活性。[结果] 从大批黄萎病抗性棉(海 7124)根际土壤中筛选出编号为YIN9的酵母菌菌株,分类鉴定结果表明:YIN9菌株属于假丝酵母属Candida。平皿对峙实验结果表明:菌株YIN9对大丽轮枝菌的抑菌率达59%;将菌株YIN9的无菌发酵滤液与大丽轮枝菌孢子共培养12 h后镜检发现,用菌株YIN9处理的实验组,大部分棉花黄萎病病菌孢子不能正常萌发。盆栽实验结果表明:菌株YIN9对棉花黄萎病的平均防治效果为60.02%,可以显著降低感病棉棉花黄萎病的发病率和病情指数。此外,与从黄萎病抗性棉根际土壤中筛选获得的其他酵母菌株相比,菌株YIN9具备较高的杀线虫活性:菌株作用全齿复活线虫48 和60 h后,线虫死亡率分别为90%和100%。将菌株YIN9发酵液煮沸后,其抑制大丽轮枝菌和杀线虫活性均急剧下降,进一步测试发现,该菌株拥有较高的蛋白酶和纤维素酶活性。[结论] YIN9中的生防因子可能是热不稳定性物质,具备较高的杀线虫活性,可以显著提高感病棉对黄萎病的抗性。     

椰子脂肪酸积累及与FatB基因表达关系研究

为筛选控制椰子果实中低碳链脂肪酸积累的关键FatB基因,本研究以海南本地高种绿椰四个发育时期的果实为试验材料,参考国家标准GB/T2906-1982谷类、油料作物种子粗脂肪测定方法,用索氏提取法提取果肉脂肪酸,应用气质联用检测技术测定各发育时期脂肪酸含量及组分,同时应用qRT-PCR对椰子脂肪酸合成相关基因FatB1、FatB2及FatB3进行相对定量分析,明确各发育时期椰子果实脂肪酸积累与FatB基因表达量变化之间的关系。结果表明:(1)椰子油含9种脂肪酸成分,在椰子果实发育的早期(第6和第7个月),棕榈酸、硬脂酸和油酸的含量较高,月桂酸和肉豆蔻酸相对含量较低,通过实时荧光定量PCR发现,此时FatB1基因的相对表达量较高。(2)而在椰子果实发育的后期(第8和第9个月),棕榈酸、硬脂酸和油酸的相对含量迅速降低,月桂酸和肉豆蔻酸相对含量迅速升高,FatB2和FatB3基因的相对表达量较高,FatB1的表达量略有降低。该研究初步掌握了椰果各发育时期FatB基因表达量与脂肪酸成分间的变化趋势,该结果为将来筛选控制椰子果实中低碳链脂肪酸积累的关键FatB基因奠定基础,同时为椰子脂肪酸成分的分子改良提供理论依据。     

Relationships between rhizoplane and rhizosphere bacteria and verticillium wilt resistance in potato

Six cultivars and breeding lines of potato (Solanum tuberosum) differing in susceptibility to verticillium wilt caused by Verticillium dahliae were studied with respect to quantitative and qualitative differences in the bacterial flora of their soil and rhizosphere-rhizoplane. Although, no association was observed between the types of bacteria that inhabited the soil or roots of wilt resistant and susceptible cultivars, quantitative differences were evident. These differences provide the first direct evidence that potato genotypes can influence bacterial populations. Bacterial populations were 9–25-fold higher on roots than in the adjacent soil. As the plants aged, the total number of rootcolonizing bacteria increased between 15 and 245%. Pseudomonas spp. were the most abundant microbes in the soil and rhizosphere-rhizoplane. The bacteria antagonistic to V. dahliae in vitro were identified as members of the genera Bacillus, Pseudomonas, Flavobacterium, and Gluconobacter. A statistically significant trend was evident toward the association of antagonistic bacteria with the more resistant potato cultivars.     

Impact of Plant Species and Site on Rhizosphere-Associated Fungi Antagonistic to Verticillium dahliae Kleb.

Fungi with antagonistic activity toward plant pathogens play an essential role in plant growth and health. To analyze the effects of the plant species and the site on the abundance and composition of fungi with antagonistic activity toward Verticillium dahliae, fungi were isolated from oilseed rape and strawberry rhizosphere and bulk soil from three different locations in Germany over two growing seasons. A total of 4,320 microfungi screened for in vitro antagonism toward Verticillium resulted in 911 active isolates. This high proportion of fungi antagonistic toward the pathogen V. dahliae was found for bulk and rhizosphere soil at all sites. A plant- and site-dependent specificity of the composition of antagonistic morphotypes and their genotypic diversity was found. The strawberry rhizosphere was characterized by preferential occurrence of Penicillium and Paecilomyces isolates and low numbers of morphotypes (n = 31) and species (n = 13), while Monographella isolates were most frequently obtained from the rhizosphere of oilseed rape, for which higher numbers of morphotypes (n = 41) and species (n = 17) were found. Trichoderma strains displayed high diversity in all soils, but a high degree of plant specificity was shown by BOX-PCR fingerprints. The diversity of rhizosphere-associated antagonists was lower than that of antagonists in bulk soil, suggesting that some fungi were specifically enriched in each rhizosphere. A broad spectrum of new Verticillium antagonists was identified, and the implications of the data for biocontrol applications are discussed.     

Effect of the chemical and physical condition of the soil on Verticillium wilt of antirrhinum

Summary In vitro study showed thatVerticillium dahliae Kleb. grew well in a wide range of acid and alkaline media (viz. pH 3.5 to 10.5). The best growth of the fungus was observed in pH 5.5. Soil pH 3.5 was toxic for growing antirrhinum seedlings. Development of Verticillium wilt of antirrhinum was affected by soil pH. The severity of the disease was greater in alkaline soil conditions compared with acid conditions. Soil of pH 3.5 gave very good control of the symptom expression by the infected plants. Rhizosphere analysis results showed that fungal population with the exception ofPenicillium spp., was drastically reduced in the rhizosphere of the plants grown in acid soil. Although the overall population of fungi was reduced in theV. dahliae infected antirrhinum rhizosphere in acid soil, the population ofPenicillium spp. markedly increased. The antagonistic activity of thePenicillium spp. in the rhizosphere might also have reduced the disease severity. Since the seedlings did not grow properly in very dry and very wet soil, rhizosphere analysis of these soils was not possible. Disease severity was much less in wet soil compared with plants grown in medium moisture level and dry soil, but the plant growth was very poor. Dedicated to the memory of the late Prof. H. K. Baruah from whom I had the inspiration for research