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。     

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。     

豆蔻酸和棕榈酸诱导茄子根际拮抗菌与黄萎菌数量消长的关系

以气质联用仪(GC-MS)鉴定的嫁接茄与自根茄根系分泌物主要差异成分豆蔻酸和棕榈酸,利用它们的模拟物质研究了其对茄子根际土壤黄萎菌拮抗真菌与细菌数量和根际土壤黄萎菌数量的影响,明确了两模拟成分对茄子根际拮抗真菌和拮抗细菌的诱导作用及对黄萎菌数量抑制效果。同时,进一步分析了茄子根际拮抗真菌与细菌数总和与根际土壤黄萎菌数量的相关性。结果表明豆蔻酸和棕榈酸处理,茄子根际拮抗真菌和细菌数量均有所增加,在中高浓度下拮抗真菌和细菌数量增加显著。两物质处理下拮抗真菌与细菌数量在开花期和结果期增加显著。两物质对茄子根际黄萎菌数量的影响,整体来说,随两模拟组分浓度的增加土壤黄萎菌数量有所减少,各时期表现有所差异,在开花期两物质处理显著减少了黄萎菌数量。相关性分析表明豆蔻酸和棕榈酸处理下茄子根际拮抗真菌与细菌总量与根际黄萎菌数量呈显著的负相关性。说明嫁接通过改变茄子根系分泌物成分与对茄子根际土壤黄萎菌拮抗菌的诱导有一定关系。     

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。     

嫁接对茄子根系分泌物中肉桂酸和香草醛的调节效应

茄子根系分泌物中普遍存在肉桂酸和香草醛等化感物质.本文采用盆栽试验研究了嫁接对这两种物质的调节效应.结果表明：嫁接降低了茄子根系对这两种物质的分泌,尤其是显著降低了对香草醛的分泌,与自根茄子相比,嫁接使根系分泌的肉桂酸最多降低68.96%,香草醛最多降低100%.在外源肉桂酸和香草醛胁迫下,特别在外源肉桂酸胁迫下,与自根茄子相比,嫁接促进了茄子植株的株高、茎粗,使叶绿素含量增加5.26%～13.12%,降低了根系电导率和MDA含量,提高了根系的SOD活性,明显缓解了肉桂酸的胁迫.     

嫁接茄子根系分泌物的化感效应

采用室内生物测定的方法,研究了不同砧木、不同浓度和不同生育期嫁接茄子根系分泌物的化感效应.结果表明,嫁接茄根系分泌物与自根茄相比,促进了茄子种子的萌发和幼苗生长.各砧木品种嫁接茄根系分泌物的化感效应显著或极显著高于对照,其中发芽率和苗高最高,增加29.1%和37.1%;不同浓度试验结果呈现“低促高抑”的规律,当浓度为0.04g·ml^-1时发芽率最高,比对照增加50%,当浓度增加到0.24g·ml^-1时,根长比对照降低了30.3%;在生长后期嫁接茄的根系分泌物对种子萌发的促进作用小于前中期,而自根茄抑制作用增强.因此,生产中使用嫁接技术是缓解由自毒作用引起的连作障碍的有效方法之一.     

多菌灵胁迫下嫁接对茄子生长和抗氧化酶活性的影响

以野生茄托鲁巴姆(Solanum torvum)为砧木,以栽培品种西安绿茄(S.melongena)为接穗进行嫁接,测定不同施入频次多菌灵条件下嫁接茄子和自根茄子果实中的农药残留、生长发育状况和抗逆生理指标。结果表明,经过多菌灵施药1次、2次、3次处理后,嫁接茄果实中农药残留量比自根茄分别降低了12.7%～49.6%、11.4%～63.1%、13.5%～44.1%;嫁接茄株高、茎粗、植株鲜重和鲜果重相对于自根茄明显增加;嫁接茄根系活力比自根茄增加了6.8%～29.3%、8.7%～24.5%、11.1%～28.6%;嫁接茄叶片过氧化氢酶(CAT)活性、过氧化物酶(POD)活性、谷胱甘肽过氧化物酶(GSH-PX)活性相对于自根茄明显增加,而相对电导率比自根茄降低了16.3%～27.3%,19.7%～31.9%、20.5%～36.1%;嫁接茄丙二醛含量比自根茄降低了21.4%～31.6%、24.6%～37.9%、23.7%～39.0%。试验结果显示,施药2次和3次,各指标变化相对于施药1次明显。因此,在多菌灵胁迫下,嫁接茄子果实中的农药残留降低,植株长势明显增强,有关酶的代谢活性显著提高,从而缓解了多菌灵胁迫对茄子造成的危害。     

咖啡根系分泌物对嫁接后植株生长及叶片保护酶活性的影响

咖啡根系分泌物引起的自毒作用是造成咖啡连作障碍的主要原因,而种间嫁接是抑制咖啡自毒的有效方式。为探明嫁接抑制自毒机制,采用盆栽试验研究咖啡主栽品种根系分泌物对嫁接植株的化感效应。结果表明:根系分泌物抑制了咖啡自根苗生长,且随着根系分泌物浓度升高,抑制作用增强;低浓度根系分泌物能促进咖啡嫁接苗生长,经根系分泌物处理后的嫁接苗株高、根长、地上部干重和根干重分别较未处理的对照嫁接苗平均增加了22.15%、6.52%、44.80%和20.67%,根系活力、叶片保护酶活性也明显增加;高浓度根系分泌物虽未能促进嫁接苗生长,但嫁接苗较自根苗生长优势显著,经根系分泌物处理后嫁接苗POD、CAT和SOD活性分别较自根苗增加了60.61%、27.27%和18.24%,MDA含量显著减少,根系活力显著增加。因此,在根系分泌物处理下,嫁接苗能通过提高自身保护酶活性等方式,促进植株生长或使植株生长所受抑制减少。     

肉桂酸和香草醛对嫁接茄子根系生长及生理特性的影响

以“托鲁巴姆”茄子为砧木,“西安绿”茄子为接穗,采用盆栽试验研究了自毒物质肉桂酸和香草醛对嫁接、自根和砧木茄子根系生长及抗氧化酶活性、渗透调节物质含量的影响.结果表明：肉桂酸、香草醛对茄子根系生长及生理代谢的化感作用基本表现为“低促高抑”,两物质对自根茄子根系表现促进、抑制效应的临界浓度点分别为0.1和0.5 mmol·kg^-1;对嫁接、砧木茄子表现促进、抑制效应的临界浓度点分别为0.5和1 mmo·kg^-1.茄子根系对自毒物质耐性大小依次为：砧木茄>嫁接茄>自根茄;在较高浓度肉桂酸(0.5～4 mmol·kg^-1)和香草醛(1～4mmol·kg^-1)作用下,与自根茄子相比,嫁接茄子根系SOD活性提高了8.50%～24.50%,脯氨酸含量增加了9.39%～27.64%,可溶性糖含量增加了12.77%～81.81%,可溶性蛋白含量、根系鲜、干物质量和根系活力显著高于自根茄.表明嫁接换根使茄子根系具有了砧木抗自毒物质的特性,缓解了自毒物质给茄子根系生长带来的不良影响.     

镉腔迫下小麦根系分泌物的变化

通过水培和砂培两种方法,研究了镉胁迫下小麦（Triticum aestivm L.)根系分泌物变化。通过研究镉对小麦根系分泌氨基酸和糖,根系分泌其它一次次生代谢物质等的影响,以及根系分泌物对蔬菜种子萌发的影响,探讨镉胁迫下植物根系的生理生态效应。研究结果表明：镉胁迫下,小麦根系分泌物无论在量上还是质上都有变化。镉胁迫下根系分泌的电解质,糖类,氨基酸以及其它一些次生代谢物质都有所变化,但变化情况有差异。电解质外渗率,糖类随Cd^2＋浓度升高而增加,在低浓度Cd^2 作用下（0.5mg/L),随处理浓度的升高,氨基酸分泌量增加,当处理浓度高于相应浓度时,氨基酸分泌量随浓度升高而减少。随Cd^2 浓度升高,次生代谢物分泌种类减少。这说明小麦通过改变根分泌作用而缓解镉危害。由于根系分泌次生代谢物的活动比较复杂,因而根系的分泌作用有待于进一步研究。根系分泌物中可能含有某些物质抑制萝卜,白菜种子萌发,这是涉及到植物他感作用的问题,值得并有待于进一步研究。初步分析小麦根系在不同的环境中对镉胁迫的响应方式,为以后根分泌物的深入研究提供了一定的思路,也为镉污染区农作物的合理栽培提供新的参考资料。     

Use of Cucumis metuliferus as a Rootstock for Melon to Manage Meloidogyne incognita

Root-knot nematode-susceptible melons (Cantaloupe) were grown in pots with varying levels of Meloidogyne incognita and were compared to susceptible melons that were grafted onto Cucumis metuliferus or Cucurbita moschata rootstocks. In addition, the effect of using melons as transplants in nematode-infested soil was compared to direct seeding of melons in nematode-infested soil. There were no differences in shoot or root weight, or severity of root galling between transplanted and direct-seeded non-grafted susceptible melon in nematode-infested soil. Susceptible melon grafted on C. moschata rootstocks had lower root gall ratings and, at high nematode densities, higher shoot weights than non-grafted susceptible melons. However, final nematode levels were not lower on the grafted than on the non-grafted plants, and it was therefore concluded that grafting susceptible melon on to C. moschata rootstock made the plants tolerant, but not resistant, to the nematodes. Grafting susceptible melons on C. metuliferus rootstocks also reduced levels of root galling, prevented shoot weight losses, and resulted in significantly lower nematode levels at harvest. Thus, C. metuliferus may be used as a rootstock for melon to prevent both growth reduction and a strong nematode buildup in M. incognita-infested soil.     

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.     

MiR395 Overexpression Increases Eggplant Sensibility to <Emphasis Type="Italic">Verticillium dahliae</Emphasis> Infection

Verticillium wilt (V. wilt), a notorious wilt disease caused by Verticillium dahliae, often leads to the reduction of eggplant (Solanum melongena L.) production. MiRNAs, as a class of small RNAs, can regulate gene expression and then affect growth and development in plants. MiR395 has been proven to respond to sulfate-deficient stress in Arabidopsis thaliana and sulfate is well known to have a close relationship with plant disease resistance. To explore the function of eggplant miR395, we examined its expression in V. dahliae-infected eggplant by qRT-PCR and found miR395 exhibited a gradual reduction trend with time after infection. We then expressed pre-miR395 from Arabidopsis thaliana in Suqi eggplant and resistance analysis showed that miR395 overexpressed plants were hypersensitive to V. dahliae infection. We further measured the content of GSH and activities of POD and SOD and the results indicated that the index of GSH/POD/SOD in the overexpressed plants was lower than that of the wild-type control under V. dahliae infection. These results suggest that miR395 plays a negative role in eggplant response to V. dahliae infection.     

Increased resistance to fungal wilts in transgenic eggplant expressing alfalfa glucanase gene

The wilt diseases caused by Verticillium dahliae and Fusarium oxysporum are the major diseases of eggplant (Solanum melongena L.). In order to generate transgenic resistance against the wilt diseases, Agrobacterium-mediated gene transfer was performed to introduce alfalfa glucanase gene encoding an acidic glucanase into eggplant using neomycin phosphotransferase (npt-II) gene as a plant selection marker. The transgene integration into eggplant genome was confirmed by Polymerase chain reaction (PCR) and Southern blot analysis and transgene expression by the glucanase activity and western blot analysis. The selected transgenic lines were challenged with V. dahliae and F. oxysporum under in vitro and in vivo growth conditions, and transgenic lines showed enhanced resistance against the wilt-causing fungi with a delay of 5–7 days in the disease development as compared to wild-type plants.     

Identification of MiRNA from Eggplant (Solanum melongena L.) by Small RNA Deep Sequencing and Their Response to Verticillium dahliae Infection

MiRNAs are a class of non-coding small RNAs that play important roles in the regulation of gene expression. Although plant miRNAs have been extensively studied in model systems, less is known in other plants with limited genome sequence data, including eggplant (Solanum melongena L.). To identify miRNAs in eggplant and their response to Verticillium dahliae infection, a fungal pathogen for which clear understanding of infection mechanisms and effective cure methods are currently lacking, we deep-sequenced two small RNA (sRNA) libraries prepared from mock-infected and infected seedlings of eggplants. Specifically, 30,830,792 reads produced 7,716,328 unique miRNAs representing 99 known miRNA families that have been identified in other plant species. Two novel putative miRNAs were predicted with eggplant ESTs. The potential targets of the identified known and novel miRNAs were also predicted based on sequence homology search. It was observed that the length distribution of obtained sRNAs and the expression of 6 miRNA families were obviously different between the two libraries. These results provide a framework for further analysis of miRNAs and their role in regulating plant response to fungal infection and Verticillium wilt in particular.