猕猴桃溃疡病菌biovar 3群体MLVA分型技术的建立与应用

 丁香假单胞菌猕猴桃致病变种生物型3(Pseudomonas syringae pv. actinidiae biovar 3,Psa3)是猕猴桃溃疡病菌的世界流行群体,但仅在中国存在复杂的遗传多样性。开发适于Psa3群体分型的MLVA(multilocus variable-number tandem-repeat analysis)技术是探索中国Psa3起源与流行学特性的基础。本研究对7个Psa3菌株进行了全基因组测序,结合已公布的86个全基因组数据,进行比较分析发现,中国Psa3至少存在7个亚群;在各亚群间存在多态性的24个串联重复序列中,其中10个可以通过琼脂糖凝胶电泳区分开且变异指数合适,据此建立了适于Psa3的MLVA技术。采用该技术对分别来自贵州和陕西的62和9个Psa3菌株进行群体分型,分型结果与全基因组分析高度一致,证明该MLVA体系分型准确。MLVA分型结果表明:贵州主产区修文县Psa3有3个MLVA群体,其中亚群4的组内分化明显,代表最早发生的群体;而亚群 1和3的结构单一,且多在新果园发现,是新传入群体。总之,本研究建立了一套可用于Psa3群体分型的MLVA技术,将有助于解析中国各猕猴桃产区Psa3群体结构,以及探索中国Psa3的起源、传播和流行学特征。     

新型腐植酸有机肥对马铃薯茎基腐病的防效

马铃薯茎基腐病是影响马铃薯生产的重要病害,目前尚未有效的综合防控措施。试验的目的是通过新型腐植酸肥料与同等养分配方化肥的田间比较试验,探讨新型腐植酸有机肥对马铃薯茎基腐病预防效果。结果表明,5~15 g壤动FT/100 kg种薯配合腐植酸肥料对马铃薯生长发育安全,在出苗率、苗高、单株根数、最大根长、芽长分别优于配方化肥1.0~2.9个百分点、1.2~4.1 cm、1.1~5.2条、1.2~2.6 cm、0.3~1.1 cm;在单株薯重、平均单薯重、商品薯率、小区产量分别高于配方化肥66.7~394.4 g、4.6~12.1 g、4.3~11.4个百分点、12.23~83.46 kg/30.8m²;对主茎、匍匐茎、块茎基腐防效分别达78.29%~87.24%、75.62%~88.51%、72.05%~86.98%,增产效果达11.60%~79.12%。研究为开展农业技术防治马铃薯茎基腐病提供了新方法,对制定综合防治技术体系具有重要指导意义。     

猕猴桃细菌性溃疡病研究进展

笔者总结了近年来国内外学者对猕猴桃细菌性溃疡病的研究和发现，详细介绍了猕猴桃细菌性溃疡病的症状、危害和传播途径以及致病菌Pseudomonas syringae pv. actinidiae的特征、分类和起源，分析了其可能的致病机理并总结了猕猴桃的抗病性研究。基于目前对猕猴桃细菌性溃疡病的防治方法和相关研究发现，展望了猕猴桃细菌性溃疡病的研究方向和防治措施，提出采取预防致病菌传入、增强树势和药剂防治相结合的防治措施，为以后的研究和生产实践提供了理论依据。     

新疆部分地区苹果、核桃和杨树腐烂病的病原鉴定

 树木腐烂病在新疆主林果种植区均有不同程度的发生,近年呈加重趋势。本研究在调查了新疆阿克苏、和田、伊犁、喀什和库尔勒等地区苹果树、核桃树和杨树3种林果腐烂病田间发病情况的基础上,重点对14个样本点的样品进行病原物的分离与鉴定。结果表明,各样本点3种林果均有不同程度的腐烂病发生,分离鉴定结果为引起苹果树腐烂病病原菌主要为Valsa mali var. mali(分离频率为64.5%),其次是V. mali var. pyri(25.8%)、V. malicola(3.2%)和V. nivea(6.5%);引起核桃树和杨树腐烂病病原菌为V. sordida(分离频率为83.3%和96%)和V. nivea(分离频率为16.7%和4%)。离体枝条及苹果果实上的致病性测定结果表明,3种林果上分离得到的病原菌均能感染原寄主。引起新疆部分地区苹果树腐烂病的主要病原菌是Valsa mali var. mali,核桃树和杨树腐烂病的主要病原菌是V. sordida。同时,基于实际生产中存在感病枯死杨树枝干作为苹果园、核桃园树体支撑木的现象,推测3种林果腐烂病菌间可能存在交叉感染的潜在风险。     

Response of soil respiration to nitrogen addition in two subtropical forest types

Anthropogenic activities have increased nitrogen (N) deposition in terrestrial ecosystems, which directly and indirectly affects soil biogeochemical processes, including soil respiration. However, the effects of the increases in N availability on soil respiration are not fully understood. In this study, soil respiration was measured using an infrared gas analyzer system with soil chambers under four N treatments (0, 5, 15, and 30 g N m^-2 year^-1 as control, low N (LN), moderate N (MN), and high N (HN), respectively) in camphor tree and slash pine forests in subtropical China. Results showed that soil respiration rates decreased by 37% in the camphor tree forest and 27% in the slash pine forest on average on an annual base, respectively, in the N-fertilized treatments when compared with the control. No significant differences were found in the soil respiration rate among the LN, MN, and HN treatments in both forest types as these fertilized plots reached an adequate N content zone. In addition, soil microbial biomass carbon (C) content and fine root biomass declined in N-treated plots compared to the control. Our results indicated that elevated N deposition might alter the tree growth pattern, C partitioning, and microbial activity, which further affect soil C sequestration by reducing soil respiration in subtropical forests of China.     

Characterization and pathogenicity of six Cytospora strains causing stem canker of wild apple in the Tianshan Forest,China

Cytospora species are capable of causing destructive cankers of stems belonging to a wide range of woody plant species. In severe cases, cankers may lead to dieback of twigs and branches. Little is known about the Cytospora species causing canker disease of wild apple (Malus sieversii) trees in the Wild Fruit Forest Reserve in Tianshan Forest, Xinjiang Uygur Autonomous Region, China. In this study, six Cytospora isolates belonging to two species were isolated from cankerous lesions of wild apple twigs. Based on multi‐locus phylogenetic analysis using three DNA markers (ITS, tef1‐α and tub2) and morphological characterization, these isolates were identified as Cytospora mali and Cytospora parasitica. Temperature trials (15, 20, 25 and 30°C) showed that the optimal growth temperature for six isolates was 25°C. At a variety of temperatures, C. mali isolates tended to grow faster than isolates of C. parasitica, with the C. mali isolate, EGI1 performing better than others with regard to growth rate. Morphological observations showed that these species exhibited a single locule without conceptacles, and the conidia length was 3–5 μm. In vitro inoculation trials of twigs and leaves of M. sieversii seedlings revealed that the C. mali isolates were highly virulent phytopathogenic fungi, whereas the C. parasitica isolates were less virulent. The isolate EGI1 was the most virulent isolate among the six isolates. This paper presents the first report of pathogenic Cytospora spp. of the M. sieversii Tianshan Wild Fruit Forest Reserve of Yili, Xinjiang in China. It will aid in the understanding of how apple tree cankers are induced and provide disease management guidelines for M. sieversii forest conservation.     

梨CRK家族基因及其腐烂病菌侵染响应成员的鉴定

在植物响应逆境过程中，半胱氨酸富集类受体激酶（Cysteine-rich receptor like kinase，CRK）起重要的调控作用。本研究中以结构域Stress-antifung（Pfam：PF01657）和Pkinase（Pfam：PF00069）的保守序列为种子序列，在全基因组范围鉴定了梨（Pryus spp.）CRK家族成员。此外，对其蛋白理化性质、进化特征、基因在染色体上的位置、顺式作用元件（cis-acting regulatory element，cis-element）和表达模式进行了分析。共获得32个CRK家族成员，其氨基酸数、分子量和等电点分别介于440 ~ 1 217、48.90 ~ 137.02 kD和5.31 ~ 8.59，主要位于质膜。根据进化分析，将来自梨、拟南芥、苹果、番茄和水稻的156个CRK分为6个亚组，梨CRK主要分布于亚组Ⅳ，Ⅴ和Ⅵ。梨CRK中存在5个串联重复的基因簇，共包含了20个成员。此外，该基因家族所有成员的启动子区域都存在多个响应激素和逆境信号的cis-element。接种腐烂病菌Valsa pyri（Vp）后，杜梨（Pyrus betulifolia，抗病）和‘早酥’梨（Pyrus bretschneideri，感病）中分别发现8个和10个CRK发生了差异表达，6个基因在两种资源中都发生了差异表达。差异基因中，Pbr001477.1和Pbr000205.4在抗、感病资源中都显著上调，而其他基因的表达量在两种资源中呈现不同的变化趋势。     

群体感应信号物质对吡咯伯克霍尔德氏菌JK-SH007在杨树内定殖的影响

为了明确杨树溃疡病生防菌吡咯伯克霍尔德氏菌JK-SH007的群体感应系统是否与其内生定殖相关，本文通过群体感应指示菌紫色杆菌CV026和液相色谱串联四级杆飞行时间质谱技术（HPLC-Q-TOF-MS）确定其群体感应信号物质种类，并利用结晶紫染色、菌体回收、GFP标记等技术，研究其群体感应信号物质对该菌株生物膜及在杨树苗内的定殖能力的影响。结果表明，菌株JK-SH007产生的群体感应信号物质为含8个碳原子酰基侧链的辛酰基-L-高丝氨酸内酯（C₈-HSL）。群体感应信号物质C₈-HSL的添加对该菌株生物膜及在杨树苗内的定殖能力有影响，并且呈现低浓度促进，较高浓度抑制的规律。C₈-HSL的添加与菌株JK-SH007生物膜的OD值和菌体量具有显著相关性，相关系数分别为0.923和0.756。当添加1.0% C₈-HSL终浓度达到5×10^{-8 g/L}时，菌株JK-SH007生物膜的形成量达到峰值。荧光显微镜镜检发现，当添加100 μL C₈-HSL时，杨树组培苗根和茎中GFP荧光标记的菌株JK-SH007的定殖数量明显较CK多。菌株JK-SH007的群体感应与其在杨树内的内生定殖能力密切相关，群体感应信号物质C₈-HSL具有增强菌株JK-SH007内生定殖的能力，同时也表明该物质有利于该菌株生防效果的发挥。     

Azospirillum brasilense mitigates water stress imposed by a vascular disease by increasing xylem vessel area and stem hydraulic conductivity in tomato

Water stress, with its negative consequences on plant growth and survival, can be mitigated by Azospirillum brasilense inoculation. In tomato, A. brasilense delays wilting caused by a vascular pathogen, Clavibacter michiganensis subsp. michiganensis, by yet unknown mechanisms. We studied morphological, anatomical and physiological changes induced by A. brasilense in tomato that relate to water stress tolerance, which could explain the deferral in symptom expression. For this purpose, tomato seeds were treated or not with A. brasilense BNM65, and 5 weeks later plants were challenged with C. michiganensis subsp. michiganensis or mock inoculated with water. There was a large growth promotion associated to Azospirillum: treated plants had higher total biomass and leaf area. In relation to water stress tolerance, Azospirillum treated plants had larger xylem vessel area, higher stem specific hydraulic conductivity, thicker stems, and lower shoot/root dry matter and specific leaf area. These changes were opposite to those induced by C. michiganensis subsp. michiganensis. We conclude that A. brasilense favoured a better adjustment of plant-water relations by several mechanisms, and thus, transitorily alleviated symptoms expression of a vascular disease.     

合成樟脑中主要杂质的研究

本文研究了我国合成樟脑的组成,用气相色谱和气一质联用方法分析。结果表明,主要杂质是樟脑的同分异构体(小茴香酮,异小茴香酮,异莰酮)和少量醇、烃(龙脑、异龙脑、异莰烷)。内标法分析表明,樟脑的含量通常在95％以上。