玉米内生细菌-解淀粉芽孢杆菌Y19-RifM在玉米根部定殖能力的研究

采用逐步提高LB平板中利福平浓度的方法,从具有促生长和防病潜力的玉米内生细菌解淀粉芽孢杆菌Y19中筛选出获得抑菌活性不变的抗利福平(250μg/m L)突变菌株Y19-Rif M,研究Y19-Rif M在玉米根部定殖能力。结果表明,玉米出苗后25、30、35、40、45、50、55 d,从根际、根表、根内均能回收到抗性菌落。根际、根表的回收菌量保持在1×103~1×104CFU/g的水平,根内保持在1×103CFU/g的水平,灭菌土中回收到的菌落数量高于自然土中菌落数量。     

固定化植物促生菌的存活性研究

芽胞菌类的植物促生和生防菌适合用于固定化细胞颗粒的制作,其贮藏时间长,180d后菌株Tu-100和A6的含菌量分别为2.3×108个/g和1.5×108个/g,并能在根际效应的影响下,在植物根际长期有效定殖.对作物土传病害有一定的抑制作用.     

青枯雷尔氏菌在不同作物根际定殖与轮作防病

青枯病是芝麻和花生的重要病害之一。为探索合理的轮作防控模式，运用青枯雷尔氏菌（Ralstonia sola⁃ nacearum）抗利福平标记菌株JXS02-L土壤接种，测定菌株在芝麻、花生、甘薯、大豆、玉米和小葱等6种作物根际土 壤和根部的定殖与消长动态，分析了不同轮作模式对芝麻/花生青枯病发生程度的影响。结果表明：播种后3w，6种 作物根际土壤菌量均低于初始接种菌量；播种后6 w，芝麻、花生、甘薯根际土壤菌量已上升至初始接种菌量水平 （3.20 ×106~4.93×106 CFU·g-1），之后菌量持续上升，大豆、玉米根际土壤菌量则持续下降；播种后12 w，大豆、玉米根 际土壤菌量比芝麻、花生、甘薯根际土壤菌量低4个数量级，小葱植种后6 w~12 w，均未检测到病菌。播种后3 w~ 12 w，芝麻、花生和甘薯根部菌量持续上升，大豆和玉米根部菌量先升后降；至播种后12w，大豆和玉米根部菌量比 芝麻、花生和甘薯根部菌量低5个数量级，小葱根部则始终未检测到病菌。芝麻-大豆-小葱-芝麻、芝麻-大豆-玉 米-芝麻2种轮作模式芝麻青枯病病情指数比芝麻-花生-甘薯-芝麻轮作模式分别降低19.95%、12.87%；花生-大 豆-玉米-花生轮作模式花生青枯病病株率比花生-芝麻-甘薯-花生轮作模式降低11.63%。本研究结果对于了解 青枯雷尔氏菌的生态多样性，以及指导青枯病的科学防控具有重要意义。     

生防枯草芽孢杆菌Czk1固体发酵工艺条件优化

枯草芽孢杆菌(Bacillussubtilis)Czk1是一株具广谱拮抗、促生和诱导植株产生抗病性的根际益生菌。为了全面资源化利用农业废弃物中的有机养分和促进对植物土传病害的生物防治,本研究以Czk1为研究对象,以鸡粪为底肥,分别利用玉米秸秆、菌糠、椰糠堆肥为原料,采用液固两相发酵工艺,以稀释平板菌落计数法测定活菌总数和芽孢数,经过单因素和正交试验方法,明确Czk1固体发酵物料的配方、工艺参数和培养条件。得出Czk1固体发酵的最优条件：最佳原料为菌糠鸡粪堆肥,Czk1接种重量为10%,黄豆粉添加量为10%,含水量为40%,发酵温度为37℃,翻抛1次/72h,在此条件下,Czk1菌株中的芽孢活菌数量总量可以达到8.63×10⁸ CFU/g,芽孢数为8.37×10⁸ CFU/g。其次,发酵效果较佳的原料为秸秆鸡粪堆肥,Czk1接种量为10%,添加5%黄豆粉,40%含水量,发酵温度为37℃,翻抛1次/72 h,此条件下,Czk1接种菌株中的活菌数量总量可达到6.53×10⁸ CFU/g,芽孢数为5.97×10⁸     

解磷细菌 Burkholderia 的分离鉴定及对香草兰生长和 P 吸收 的影响

香草兰为喜磷作物，从香草兰种植园中分离筛选到一株解磷微生物——伯克霍尔德氏菌 V-29。在 NBRIP 液体 培养基中摇床振荡培养 5 d 后可溶性磷含量达 475.3 μg/mL，培养基 pH 下降。通过大田试验研究了接种绿色荧光蛋白 标记后的解磷菌株 V-29 及其与有机肥发酵制得的微生物有机肥对香草兰生长和磷素吸收的影响。结果表明：单独接种 V-29 或施用微生物有机肥可显著增加香草兰植株干重、土壤有效磷含量；移栽 4 个月后，标记菌株 V-29 在香草兰根际 土壤中的含量可达 106 cfu/g 土壤。由此可见，伯克霍尔德氏菌 V-29 可单独作为生物菌剂或与有机肥发酵制得微生物有 机肥后用于农业生产中，以减少化肥施用量。     

解磷细菌Burkholderia的分离鉴定及对香草兰生长和P吸收的影响（英文）

香草兰为喜磷作物,从香草兰种植园中分离筛选到一株解磷微生物——伯克霍尔德氏菌V-29。在NBRIP液体培养基中摇床振荡培养5d后可溶性磷含量达475.3μg/mL,培养基pH下降。通过大田试验研究了接种绿色荧光蛋白标记后的解磷菌株V-29及其与有机肥发酵制得的微生物有机肥对香草兰生长和磷素吸收的影响。结果表明:单独接种V-29或施用微生物有机肥可显著增加香草兰植株干重、土壤有效磷含量;移栽4个月后,标记菌株V-29在香草兰根际土壤中的含量可达106 cfu/g土壤。由此可见,伯克霍尔德氏菌V-29可单独作为生物菌剂或与有机肥发酵制得微生物有机肥后用于农业生产中,以减少化肥施用量。     

拮抗内生细菌BEB17对4个香蕉品种组培杯苗的定殖、促生分析和抗病性评价

BEB17是分离自健康香蕉根部的具有良好抑菌活性的内生解淀粉芽孢杆菌(Bacillus amyloliquefaciens)。本研究采用直接灌根法,评价了该菌株对4个不同香蕉品种组培杯苗的促生作用和其对香蕉枯萎病菌(Fusarium.oxysporum f.sp.cubense race 4,Foc4)的防治效果。结果表明,经BEB17回接后的香蕉杯苗平均株高、叶片数和鲜重均显著优于对照,其中对南天黄品种的促生效果最好,株高增加48.24%,鲜重增加111.19%;带菌植株接种Foc4后,香蕉杯苗在发病率和病情指数上均显著低于对照,其中该菌株对南天黄和桂蕉1号的防治效果最明显,相对防效分别可达81.51%和77.46%。通过稀释平板涂布法,观察其在不同品种香蕉苗的定殖情况,发现BEB17能稳定定殖于4种香蕉品种,其中在南天黄的根部定殖10 d后定殖量最高,达6.43×10~4 CFU/g,在桂蕉1号定殖10 d和20 d后茎部和叶部定殖量达峰值,分别为1.58×10~3 CFU/g和1.43×10~3 CFU/g;品种比对发现菌株BEB17与南天黄和桂蕉1号的亲和性较强,定殖数量呈先升后降趋势。     

降香黄檀-檀香根际土壤高效解磷细菌的分离筛选与鉴定

利用PKO无机磷培养基从不同林龄的降香黄檀-檀香根际土壤中分离筛选高效解磷菌,为进一步研制专用生物复合肥提供依据。通过溶磷圈法筛选溶磷较强(D/d值大于1.5)的菌株19株,采用钼锑钪比色法测定其解磷能力。结果显示:菌株无机磷溶量在9.23~223.27μg/m L之间,3株菌无机磷溶量在170μg/m L以上;菌株有机磷溶量在0.43~25.05μg/m L之间,有机磷溶量在15μg/m L以上有4株。采用Salkowski比色法测定菌株分泌IAA能力,10株菌具有分泌能力,其中Dosa P25分泌IAA量达32.14μg/m L。综上所述,菌株Dosa P15、Dosa P25具有较强的解磷、分泌IAA能力。基于形态学、生理生化特性及16 S r DNA序列比较分析,初步鉴定Dosa P15属于Bacillus pumilus、Dosa P25属于Bacillus subtilis。试验同时表明,菌株溶磷量、分泌IAA量与培养液p H值之间不存在显著相关性。     

枯草芽胞杆菌在油菜根茎叶的定殖动态和生防作用研究

采用常规方法跟踪枯草芽胞杆菌Tu-100在缩影系统油菜根际的定殖情况。Tu-100在油菜不同根段部位的定殖密度从上到下呈现了逐渐递减的规律。随着接种后时间的延长而逐渐下降。在根段8cm以外的根区几乎检测不到接种菌。在油菜播种后3d,定殖密度可达最高水平（8.3×105 个/g）,然后急速下降,30d后保持相对稳定的较低水平（1.3×102 个/g）。 在油菜三片真叶时喷雾接种一次,20d后,在茎和叶上不能检测到所接种的Tu-100,并且抗菌核病的能力也逐渐减弱。     

枯草芽孢杆菌T122F菌剂对香蕉的生物效应研究

采用室内与田间试验相结合的方法研究了枯草芽孢杆菌T122F菌剂对香蕉生物效应的影响。结果表明:菌剂能明显促进香蕉生长,主要表现为香蕉株高、假茎周长、叶片数以及叶绿素含量的增加。菌剂的施用还能显著提高香蕉内生细菌数量、叶片耐菌能力和根际土壤细菌数量。在300倍液处理下,从香蕉苗根部分离到的菌量为299.75×103 cfu/g FW,是对照的7.16倍,而从香蕉根际土壤中分离到的菌量则是对照的2.20倍。此外,经菌剂处理的香蕉根际土壤水浸液对香蕉枯萎病菌孢子萌发也表现出明显的抑制作用。     

Two Bacillus amyloliquefaciens strains isolated using the competitive tomato root enrichment method and their effects on suppressing Ralstonia solanacearum and promoting tomato plant growth

Two tomato root colonizing strains, Bacillus amyloliquefacien CM-2 and T-5, were isolated after the enrichment procedure on the roots of tomato seedlings, and evaluated for their antagonistic activities against pathogenic Ralstonia solanacearum (RS) in vitro. Three inoculation methods were used to test their biocontrol efficacy and growth promotion effects in greenhouse based on their colonization ability in rhizosphere soil. Both CM-2 and T-5 strains showed strong biocontrol and growth promotion effects on tomato seedlings. The best biocontrol efficacy was obtained by treating both seedlings and soils with the biocontrol agents. In comparison to the control, the disease incidence was reduced by 70.1 and 79.4% for CM-2 and T-5, respectively. The numbers of colony-forming units of RS in rhizosphere soil were significantly (P < 0.05) decreased as compared to the control. The density of both strains in the rhizosphere soils remained at a high level (≥10⁷ CFU/g) during a five-week period. Both strains were recovered from the interior of the stems and roots of plants. They showed positive reactions for ammoina, indole acetic acid and siderophores production, and phosphate solubilizing activity. Our data proved the potential of isolated strains for the biocontrol of tomato bacterial wilt.     

利用少孢节丛孢(Arthrobotrys oligospora)HNQ11菌株控制象耳豆根结线虫危害

象耳豆根结线虫(Meloidogyne enterolobii Yang and Eisenback)是海南岛独有的一种致病力很强的根结线虫。少孢节丛孢(Arthrobotrys oligospora Fresenius)HNQ11菌株是从海南岛香蕉(Musa sapientum Linn.)根围土壤分离到的1株捕食线虫真菌,本实验研究了其对象耳豆根结线虫的控制作用。结果表明,离体条件下HNQ11菌株能迅速产生捕食网和捕食环等捕食器官来捕食象耳豆根结线虫J2。其捕食率在J2加入HNQ11培养平板96h后即达83.5%;在盆钵控病实验中,每株番茄(Lycopersicon esculentum Mill.)幼苗用2g菌丝处理根部,根结形成比对照减少50%,根系鲜重增加147.3%。同时,在菌丝处理33d后还能在显微镜下观察到根片段上附着的HNQ11菌丝和分生孢子,重新分离菌丝和分生孢子还能在培养基上生长,说明HNQ11菌株在番茄根部具有定植能力。     

Population dynamics of Xanthomonas campestris pv. vitians on different plant species and management of bacterial leaf spot of lettuce under greenhouse conditions

The population dynamics of Xanthomonas campestris pv. vitians (Xcv) was studied both externally and internally in lettuce, tomato and pepper plants. In addition, the application of bactericides during transplant production period was carried out for the management of bacterial leaf spot of lettuce under greenhouse conditions. Epiphytic populations of Xcv were recovered on leaves of lettuce plants (10⁵ CFU/g) 5 weeks after sprayed than the other plant species when inoculated with 10⁸ CFU/ml of Xcv. When plants of each crop species infiltrated with the bacterium at 10⁵ CFU/ml, the highest populations were developed in lettuce (10⁸ CFU/cm²) followed by pepper with 10⁶ CFU/cm² and tomato with 10⁵ CFU/cm² 10-days after infiltration. Application of a mixture of Maneb and Kocide or Kocide alone as a foliar spray on lettuce significantly reduced the incidence and disease severity of bacterial leaf spot by 29 and 27% respectively. Spread of the bacterium and development of the disease may partly be managed by avoiding intercropping of these plants commonly grown in close proximity to lettuce. For the management of leaf-associated populations of Xcv in lettuce, use of a mixture of Maneb and Kocide is advocated to minimize the effect of attacks.     

Effect of time between soil infestation with Pochonia chlamidosporia and planting on the efficacy of the fungus in managing Meloidogyne javanica

The fungus Pochonia chlamydosporia acts on populations of the root-knot nematode by parasitizing their eggs before they hatch, preventing the release of juveniles. In this study, the effect of the time at which the fungus is applied to soil infested with the nematode, before transplanting tomato and lettuce seedlings, was evaluated. The reduction of symptoms and the nematode population in the host plant was also evaluated, as well as the fluctuation of the fungus population during the studied period. The seedlings were transplanted at 0, 5, 10, 15 or 20 days after incorporation of the inocula of the fungus and the nematode into the soil. The population of P. chlamydosporia, without the presence of plants, increased and reached its maximum 10 days after its incorporation into the soil. At the end of the experiments, the number of colony forming units (CFU) of the fungus in the soil remained high enough to act on the next cycle of crops. For both cultures, the greatest effect on reducing the number of galls and eggs of Meloidogyne javanica was obtained by increasing the contact time of the fungus with the nematode in the soil before transplanting seedlings. In the second run, for both cultures, there was an increase in shoot weight with increasing of incubation time before the transplantation of seedlings.     

Maximizing rhizosphere populations of fluorescent pseudomonads on potatoes and their effects onErwinia carotovora

Field-grown potato plants were treated withPseudomonas putida andP. fluorescens strains in order to maximize the rhizosphere populations of the pseudomonads and determine their effects on indigenous populations ofErwinia carotovora. Single applications of the pseudomonads to seed pieces at planting resulted in peak populations of approximately 10⁵ colony-forming units (CFU) per gram (fresh weight) of root in July, whereas a second application in June boosted rhizosphere populations 10- to 100-fold higher throughout the remainder of the growing season; they composed nearly 20% of the average total population of rhizosphere bacteria in early July. Although rhizosphere populations of the introduced pseudomonads were high,E. carotovora populations sharply increased from very low numbers to reach populations ranging between 10⁵ and 10⁷ CFU/g by early July. Furthermore, coapplications of pseudomonad strains failed to suppress native populations ofE. carotovora despite the growth and maintenance of high rhizosphere populations of both pseudomonad strains.     

腐霉利在番茄上的残留分析

为了评价腐霉利在番茄上的残留状况,指导腐霉利的科学合理使用,本试验选择50%腐霉利可湿性粉剂推荐剂量500.0、750.0 g/hm²(有效成分),连续施药2～3次,间隔5 d,最后一次施药3、7、10 d后采集番茄样品进行残留检测。结果表明,腐霉利在番茄上的残留量为0.075～0.256 mg/kg,小于GB 2763-2012规定的农药最高残留量(2 mg/kg);结合番茄成熟期长,建议腐霉利在番茄上的安全间隔期为10 d。     

解淀粉芽孢杆菌HAB-7对18株植物病原真菌的抑制作用

HAB-7是一株分离自豇豆根际土壤、具有较强抑菌作用的解淀粉芽孢杆菌(Bacillus amyloliquefaciens)。研究HAB-7对18株植物病原真菌及其发酵液粗提蛋白对植物病原的抑制作用，并测定其对番茄种子的促生作用。采用平板对峙法测定其对18株植物病原真菌的抑制活性，以橡胶树炭疽菌为指示菌，对其发酵上清液经60%、80%硫酸铵盐析获得的抗菌粗蛋白进行测试；并将HAB-7菌液稀释成6个浓度梯度，分别用来对番茄种子进行促生测试。结果显示：HAB-7对18株植物病原真菌的平均抑菌率为61.03%；     

桐花树内生细菌动态及拮抗植物病原菌菌株筛选鉴定

以荧光显微计数法和平板分离法分别对桐花树根茎叶内生细菌总量和可培养细菌数量进行周年动态初步测定,并对拮抗植物病原菌的可培养细菌菌株进行筛选鉴定。结果表明:桐花树根茎叶内均含有大量的内生细菌,其中根部内生细菌总量为0.88×107~49.67×107ind./g FW,7月份最大,叶部细菌总量为1.07×107~65.07×107ind./g FW,也以7月份最大,茎部细菌总量为1.1×107~19.73×107ind./g FW,以5月份最大;而根茎叶可培养细菌含量分别为1.02×103~13.18×103、0.13×103~11.24×103、0.31×103~10.36×103CFU/g FW,均以3月份数量最大,周年不同月份之间细菌总量和可培养细菌数量均差异显著。用平板对峙生长法,从86株可培养内生细菌中筛选获得1株对香蕉叶鞘腐败病菌、香蕉枯萎病菌、马拉巴栗茎腐病等植物病原菌均有较强拮抗作用的Aec23菌株,经形态、生理生化测试及16S rDNA序列比较分析,Aec23菌株初步鉴定为解淀粉芽孢杆菌(Bacillus amyloliquefaciens)。     

文心兰侧芽诱导丛生芽技术研究

以文心兰侧芽为外植体,研究侧芽采前处理、消毒次数、培养基中不同椰子汁含量配比对丛生芽诱导的影响。结果表明,采用在培养基中添加天然植物体,以幼嫩的椰子汁本身所含的微量激素和植物细胞分裂所需的关键诱导因子代替人工合成激素,对以文心兰侧芽为外植体进行丛生芽的诱导效果明显;以文心兰侧芽为外植体,通过采前5～7 d对植株的控水和喷施抗菌剂相结合的方法处理后,可避免病菌感染;经过连续2次0.15% 的升汞溶液灭菌(其中消毒时间为第1次7～8 min,第2次30～50 s),消毒效果最好;以(1/3大量元素)MS＋15