Validation and use of a qPCR protocol to quantify the spread of Ralstonia solanacearum in susceptible and resistant eucalypt plants

Bacterial wilt caused by Ralstonia solanacearum is a serious disease of eucalypt in humid and high temperature areas worldwide. Spreading of the bacterium in the field or to other nurseries occurs mainly by symptomless infected plant material. The use of pathogen-free propagating material as well as planting of resistant genotypes are currently the only strategies used for disease control. Therefore, a reliable and sensitive method for detection of low titres of R. solanacearum in infected plant tissue is essential for the success of management programmes. In this work, we adapted an efficient intercalating dye-based real-time PCR protocol to detect the bacterium in symptomless eucalypt plants as well as to investigate its movement in eucalypt clones CLR172 and CLR371, which exhibit resistant and susceptible phenotypes, respectively. We found that the bacterium translocates acropetally and basipetally in inoculated but symptomless cuttings of the resistant clone, as in cuttings of the susceptible clone displaying symptoms. Nevertheless, a smaller concentration of bacterial DNA was detected in tissues of the resistant clone. Mature biofilms occluding the xylem vessels were present in the susceptible clone whereas only single cells or small aggregates were observed in the resistant clone. This work contributes to improve our knowledge of the colonization process of R. solanacearum in eucalypt clones with different levels of susceptibility and to understand how the defence mechanisms against bacterial wilt in Eucalyptus work. Our findings could aid in the selection of the most resistant eucalypt clones to be used in wilt disease management programmes.     

莠去津土壤残留生物修复菌肥作用原理与田间应用

莠去津是玉米田应用的优秀除草剂品种,然而由于其在土壤中残留时间长,常对轮作后茬敏感作物造成严重毒害。采用生物修复菌肥做种肥、结合菌肥拌种和叶面喷施方法,研究对玉米后茬旱直播水稻生长发育及药害修复机理,对土壤中莠去津残留量、水稻生长和生理指标、土壤酶活性进行测定。结果表明:颗粒菌肥做种肥+粉剂菌肥拌种+水剂菌肥叶面喷施是莠去津土壤残留毒害修复的最佳方法,土壤中莠去津含量在喷施菌肥后7 d从施用菌肥前的0.9 mg/kg下降到0.1 mg/kg,水稻叶片叶绿素含量显著增加33.74%,超氧化物歧化酶和过氧化物酶酶活性分别显著提高23.39%和92.57%,丙二醛含量则显著降低48.01%;水稻株高、地上部鲜重、干重分别比对照显著增加22.33%、67.51%和74.80%,根系鲜重和干重分别比对照显著增加33.98%和55.43%;土壤磷酸酶、脲酶及纤维素酶含量分别显著增加49.17%、528.65%和35.21%。     

种稻年限对盐渍化土壤理化性质和细菌群落多样性的影响

为了解盐渍化土壤长期种稻引起土壤微生物和理化性质的变化特征,选取了种植水稻0、1、6、12、21 a的田块,用旱地(种植玉米)土壤作对照,利用Illumina Hiseq高通量测序技术解析不同种稻年限土壤细菌群落结构的特征,并分析了土壤理化因子与细菌群落结构的关系。结果表明:变形菌门、拟杆菌门、放线菌门是所有样点土壤细菌群落的优势种类,其中,变形菌门占总量的36.39%～53.20%,γ-变形菌纲相对丰度在0 a种稻的土壤中显著高于其他样点(P0.05)。变形菌门相对丰度在种稻1 a的土壤中含量较高,达到45.26%;拟杆菌门的相对丰度在种稻12 a和21 a土壤中较高,分别为20.25%和20.44%;放线菌门相对丰度在0 a土壤中显著高于其他样点(P0.05),为17.90%。在属水平,布赫纳氏菌属是所有样点的优势属。α多样性指数结果显示:种稻12 a的土壤细菌群落多样性和丰富度都显著低于其他样点,分别低于对照17.06%、85.73%、82.88%。土壤理化性质测定结果表明:全氮和硝态氮在旱地土壤中含量最高;微生物碳和电导率在种稻12 a的土壤中最低,低于对照101.89%、23.25%;有机碳在21 a种稻土壤中含量最高,高于对照29.03%;pH值在21 a种稻土壤中最低,低于对照组1.05%。Pearson相关性分析表明:微生物量碳和土壤电导率对细菌多样性指数的影响显著。冗余分析结果表明,土壤电导率值、微生物碳和硝态氮是影响盐渍化稻作土壤细菌群落结构的主要因子。研究表明,随着种稻年限的延长土壤细菌群落和理化性质发生显著变化,土壤生境呈明显的改善趋势。     

马铃薯细菌性软腐病研究进展

细菌性软腐病是一种可引起果蔬严重危害的世界性植物病害,其频发已成为制约马铃薯种植业发展的瓶颈。本文基于国内外细菌性软腐病最新的研究报道,对马铃薯细菌性软腐病的病原鉴定、发病症状、致病机理、致病因子及防治措施等进行综述,并提出了进一步的研究方向,以期为马铃薯细菌性软腐病的相关研究提供科学参考。     

广西天桃木土壤细菌群落多样性特征

2020年6月,以广西南宁市、崇左市扶绥县、百色市田阳区的原生天桃木Mangifera persiciformis为研究对象,采用PacBio第三代高通量测序技术,对天桃木土壤细菌群落多样性进行了研究,并通过测定土壤化学性质,分析天桃木土壤优势细菌群落与土壤化学因子的关系.结果表明:在门水平上,共存在23门细菌,其中变...     

晾晒、添加物及不同温度对象草青贮发酵品质和微生物的影响

华南地区气候潮湿多雨,象草收获时水分含量高,单独青贮发酵品质较差。通过晾晒（水分含量由83.19%降至75.11%）、添加10%玉米粉或2%蔗糖,探究象草在不同温度（20、30和40 ℃）下青贮的发酵特性和微生物组成。研究结果表明：温度对象草各处理的青贮饲料的发酵品质有显著影响。晾晒后原材料的水溶性碳水化合物含量显著降低,青贮后发酵品质变差,尤其在30 ℃,pH达到5.85,乳酸含量只有1.56% DM,乙酸含量为2.10% DM,NH₃-N含量高达25.09% TN。对照和添加物处理的青贮饲料均随青贮温度升高,乙酸含量增加。晾晒后在20和30 ℃时青贮,产气量显著高于40 ℃或未晾晒处理。此外,象草青贮前的细菌多样性较丰富,青贮后主要是肠杆菌属和乳球菌属,尤其是20 ℃青贮的相对丰度较高,且晾晒处理在20和30 ℃青贮肠球菌属的相对丰度高于40 ℃。因此,在华南地区,象草原料中添加玉米粉或蔗糖可以降低环境温度对象草青贮发酵品质的不良影响,阴湿条件下晾晒未发挥积极效果。     

Adenovirus precipitating antibodies in the sera of brush-tailed possums in New Zealand

Sera collected from the Australian brush-tailed possum Trichosurus vulpeculu in New Zealand in 1975 and 1989 were tested for the presence of antibodies to adenovirus. Of the 231 sera tested in an agar gel diffusion test, eight (3.5%) had precipitating antibodies to the group specific antigen of mammalian adenoviruses. Available data allowed 99/231 sera to be classified as being obtained from either adult (total 62) or sexually immature (total 37) possums. From the adult animals, 4/62 (6.5%) sera were positive, while no reactive sera were detected among the 37 immature animals. The results provide evidence that natural infection by an adenovirus occurs in possums in New Zealand. Further work needs to be carried out to isolate this adenovirus to enable detailed serological and epidemiological studies to be performed. A species-specific “possum adenovirus” could have potential in the biological control of this species.     

Comparison of three markers for the determination of bacterial protein in terminal ileal digesta in the growing pig

The aim of the study was to compare three methods commonly used to determine the concentrations of bacterial protein in digesta collected from the terminal ileum of growing pigs that had been fed a casein‐based diet. The amounts of bacterial protein in terminal ileal digesta were determined using three different markers: 2.6‐diaminopimelic acid (DAPA) and the d ‐amino acids, d ‐aspartic acid (d ‐Asp) and d ‐glutamic acid (d ‐Glu). The effectiveness of each marker was compared against a control based on physical fractionation by centrifugation. The total bacterial protein concentrations derived from the markers d ‐Asp and d ‐Glu were significantly different (p = 0.05) to those calculated from DAPA and the control, but there was no difference between DAPA and the control. The percentage of bacterial nitrogen ranged from 40% to 52% dependent on the marker used. Bacterial protein expressed as a percentage of the total protein, ranged from 48% to 62%, a substantial proportion of which (12–28%) was derived from lysed bacterial cells. Statistical correlations between the estimation methods were low. Such poor correlation between the markers may be the result of random errors such as variance in the epimerization of the two d ‐amino acids during protein hydrolysis. DAPA was accepted as a reliable marker for determining microbial protein in ileal digesta.     

Improvement of Bacterial Blight Resistance in Rice Cultivars Jyothi and IR50 via Marker-Assisted Backcross Breeding

Abstract

In pathogen population analysis of 208 Xanthomonas oryzae pv. oryzae(Xoo) strains that were assembled from different parts of India, 21 pathotypes were identified on the basis of disease reactions on near-isogenic lines (NILs) and 13 pathotypes, on rice differentials. Rice cultivars, Jyothi and IR50, which are high yielding but highly prone to bacterial blight (BB) caused by pathogen populations of Xanthomonas oryzae pv. oryzae in India, were chosen. To improve the BB resistance of these two varieties, a pyramid line, NH56, containing four R-genes, Xa4, xa5, xa13, and Xa21, was selected as the R-donor based on resistance to existing pathogen population. The four R-genes were successfully transferred to cultivars through a traditional backcross method and their presence was documented with marker-aided selection (MAS). Thirty BC₄F₂ plants derived from JxNH56 (cv. Jyothi) and 45 BC₄F₂ plants derived from IR50xNH56 (cv. IR50) had all four resistance genes (Xa4, xa5, xa13, and Xa21), which should be useful resistance donors for breeding other BB-resistant elite indica varieties.     

嫁接对辣椒次生代谢的影响及其与青枯病抗性的关系

【目的】探讨嫁接辣椒酚酸类物质代谢在提高青枯病抗性中的作用。【方法】以‘富根卫士’辣椒为砧木,以‘新丰2号’为接穗嫁接,自根嫁接辣椒为对照（CK）,通过人工接种青枯假单胞杆菌（Ralstonia sdanacearum E. F. Smith）研究嫁接与自根辣椒电解质渗漏率（EL）和过氧化氢（H2O2）含量、酚酸类物质及其相关酶活性的变化。【结果】随着接种后时间的延长,嫁接与自根辣椒的EL快速增加,但嫁接辣椒的EL升高幅度显著小于自根的;接种初期,嫁接辣椒叶片与根系中的H2O2含量显著高于自根的,而6—8 d后显著低于自根的。与自根辣椒相比,嫁接辣椒的香草醛、水杨酸、木质素含量及苯丙氨酸解氨酶（PAL）和多酚氧化酶（PPO）活性均较高。【结论】嫁接辣椒次生代谢增强,酚酸类物质增多是其抗病性增强的重要原因,水杨酸和香草醛在提高嫁接辣椒青枯病抗性中发挥主要作用。