云南甘蔗白条病的多基因联合鉴定

甘蔗白条病是由白条黄单胞菌Xanthomonas albilineans(Ashby) Dowson引起的甘蔗重要病害。2017年在云南保山、孟定和金平蔗区的甘蔗上发现疑似白条病的病害。从病株蔗茎中分离得到圆形,凸面,光滑,有光泽,黄色的细菌菌落。经柯赫氏法则验证,分离得到的细菌为该病害的病原菌。使用16S rRNA基因、gyrB基因和rpoD基因对分离得到的病原菌进行了分子鉴定,BLAST和系统发育分析表明本研究所获得的病原菌的3个基因的核苷酸序列与白条黄单胞菌GPE PC73菌株(GenBank登录号:FP565176)对应基因的核苷酸序列一致性均为100%,在系统发育树中处于同一分支。根据田间症状诊断、柯赫氏法则验证及分子鉴定结果,确认云南保山、孟定和金平发生的甘蔗病害为X.albilineans引起的甘蔗白条病。     

广西宜州甘蔗白条病调查及分子检测

甘蔗白条病是一种检疫性细菌病害,具有危害性大,潜伏期长的特点,由白条黄单胞菌（Xanthomonas albilineans）引起,在种植甘蔗的主要国家或地区均有发生。2020年在广西宜州蔗区发现疑似感染甘蔗白条病蔗株,为明确病原及发生情况,为其科学防控提供科学依据。本研究对不同甘蔗品种新植和宿根蔗进行发病率调查,并采集病样进行PCR检测分析。田间调查结果显示：不同品种自然发病率不同,桂糖44新植和宿根蔗均有白条病发生,且宿根蔗发病率高于新植蔗,其中新植蔗发病率为8%～18%,平均发病率为13.83%;宿根蔗发病率为18%～34%,平均发病率为24.71%。而桂糖42、桂糖55、柳城05-136等3个品种新植和宿根蔗均未发现白条病。PCR检测结果表明：有10份样品扩增出608 bp的特异性条带,阳性检出率为90.9%,所得序列与X. albilineans序列同源性高达100%,且在构建的系统发育树处于同一分支。本研究结果表明,广西宜州蔗区发现由X. albilineans引起的甘蔗白条病。     

广西蔗区甘蔗白条病菌的鉴定与致病力分析

甘蔗白条病是一种检疫性病害,在我国多个蔗区均有报道,严重威胁我国甘蔗生产.本研究对采自广西蔗区不同品种(系)的病原菌进行分离、鉴定、保存,并选取其中40份白条黄单胞菌分离物进行遗传多样性分析与接种试验,评价其致病性.根据三磷酸腺苷结合盒转运蛋白等管家基因和Ⅳ型分泌系统virB基因的序列差异和进化树分析,将40个菌株分为...     

宿根矮化病菌对甘蔗品质及茎、叶超微结构的影响

 甘蔗宿根矮化病(Ratoon Stunting Disease, RSD)是由Leifsonia xyli subsp.xyli (Lxx)引起的,是目前世界所有植蔗地区危害性极大的病害之一。本实验以甘蔗品种新台糖22号(ROC22)健康植株为对照,感染RSD植株为处理,观察和测定RSD侵染甘蔗引起的蔗株农艺性状、蔗糖分以及茎、叶超微结构的变化。结果表明:(1)感染RSD种茎的出苗率比对照减少2.94个百分点;株高比对照下降28.85 cm;茎径比对照减少0.28 cm;节间长度比对照减短3.50 cm;单茎重比对照低0.36 kg。(2)感染RSD植株的蔗糖分低于对照0.9个百分点(绝对值)。(3)利用透射电镜技术对感染RSD植株茎、叶细胞超微结构进行观察表明,叶片叶肉细胞、维管束鞘细胞及茎细胞内的细胞器及细胞核都发生了明显的病理变化。与健康叶片相比,叶绿体变形,叶绿体基质片层大部分消解,基粒结构消失,叶绿体外膜和内膜剥离。线粒体形态异常,有的肿大、内嵴模糊,严重者内嵴消失并空泡化,仅剩未被消解的残骸;细胞核形态变为不规则,核膜破裂,染色质分布不均匀,呈降解状态。在感染RSD甘蔗茎维管束导管细胞内积累有大量的电子致密物质,细胞壁有不同程度的溶解和断裂,这可能和RSD病原细菌侵染有关。以上结果表明:RSD侵染甘蔗后,可能导致光合效率下降,对水分和营养物质的运输能力降低,从而导致甘蔗品质和产量的降低。     

甘蔗白条病及其致病菌Xanthomonas albilineans研究进展

由白条黄单胞杆菌Xanthomonas albilineans引起的甘蔗白条病是一种寄生在植物维管组织的系统性细菌病害,在全球多数种植甘蔗的国家或地区普遍发生,对甘蔗产业的发展构成潜在威胁。本研究综述了甘蔗白条病的发生和分布、传播与流行规律以及病原菌生物学与基因组特性、鉴定与检测、遗传多样性和致病机理等方面内容;提出加强抗病种质挖掘与新品种选育推广、加快抗病分子育种进程、切断病害传播途径、加强隔离检验检疫等病害防控策略;此外,评估了该病害在我国蔗区流行暴发的风险,展望今后甘蔗抗病分子育种、病原菌致病机制及其与寄主互作机理等方面的分子基础研究重点与应用前景。     

球毛壳ND35菌株在宿主植物上的侵染定殖

为了解球毛壳Chaetomium globosum ND35菌株在宿主植物上的侵染定殖方式和途径,以毛白杨组培苗为宿主植物,借助光学显微镜、扫描电镜、透射电镜,结合免疫荧光标记技术,研究了球毛壳ND35菌株子囊孢子萌发后在毛白杨上的侵染行为及其菌丝在组培苗根部的定殖。结果显示,子囊孢子萌发后形成的菌丝,能从杨树苗根、茎部表面细胞间的缝隙侵入或在根表面形成附着胞,进而形成侵染钉直接从表皮细胞侵入,在叶部主要从气孔侵入叶片内部。侵入根部的菌丝主要定殖于表皮细胞、外皮层细胞和细胞间隙,未进入内皮层和维管束组织。     

粤北地区果蔗细菌性病害PCR分子检测

为了解粤北地区果蔗主产区甘蔗宿根矮化病、赤条病和白条病病害发生和为害,本研究通过PCR技术对粤北果蔗产地植株蔗茎样品进行分子检测。结果表明,果蔗宿根矮化病菌总检出率为41.5%,各采样地块均检测出宿根矮化病病原菌,地块检出率达到100%,不同地块的检出率为10%~82.8%。赤条病和白条病病原菌未检出。粤北地区果蔗细菌性病害以甘蔗宿根矮化病为主。     

广西主要蔗区螟虫种类调查与分析

甘蔗螟虫是甘蔗生产中的重要害虫?为掌握目前广西主要蔗区苗期螟虫的主要种类和发生情况, 本研究于2019年4月-5月, 对广西主要蔗区苗期螟害种类及其为害率进行了田间实地调查, 并分析了螟害种类结构的变化?结果表明:在宿根蔗地, 甘蔗苗期黄螟为害最严重, 造成的枯心苗率为5.59%; 其次为条螟, 造成花叶苗率为5.29%; 黄螟为害造成的宿根蔗枯心苗率以桂西南和桂南两大蔗区相对较高, 其平均值分别为7.34%和6.76%; 条螟为害造成的宿根蔗花叶苗率以桂西南蔗区最高, 平均值达7.82%, 桂中和桂南蔗区其次, 其平均值分别为4.59%和4.46%?同一蔗区, 黄螟和条螟在宿根蔗地发生均比新植蔗地严重, 在宿根蔗地平均枯心苗率分别为7.13%和7.99%, 而在新植蔗地分别为2.31%和3.43%?以黄螟枯心苗率为防治指标的达标防治蔗地样点数占比为52.06%, 条螟花叶苗率为防治指标的达标防治蔗地样点数占比为56.16%?广西蔗区甘蔗苗期的主要螟虫为黄螟和条螟, 尤其是黄螟已上升为主要害虫, 表明广西蔗区的优势螟虫种类结构已发生明显变化; 广西南部和西南蔗区的苗期螟害重于北部蔗区; 宿根甘蔗地的螟害重于新植蔗地, 宿根蔗地是螟害防治的重点?     

云南蔗区甘蔗蛀茎象近年发生趋重

云南蔗区甘蔗蛀茎象近年发生趋重甘蔗蛀茎象是云南蔗区近年发展起来的一类严重蛀食蔗茎的新害虫。自１９９２年首次在西双版纳发现以来,连年猖獗发生,为害严重。为此,我们在省科委应用基础研究基金资助下,于１９９５～１９９７年对其发生种类、分布范围及为害特点进行...     

西瓜枯萎病菌侵染西瓜根茎部的组织病理学观察

为探明西瓜枯萎病菌侵染西瓜根系及茎的过程,本研究选取一株致病力较强的西瓜枯萎病菌FO1022,利用绿色荧光蛋白基因（GFP）标记该菌株,观察病原菌侵染西瓜根茎部的组织学过程。结果表明,用GFP标记的病菌接种感病西瓜品种‘苏蜜1号’,接种后2 d,显微观察显示病原菌的分生孢子附着于西瓜根系表皮细胞并萌发,沿细胞间层定殖生长;接种后5 d,在西瓜根部维管束导管中观察到大量菌丝和大型分生孢子,在根部表皮细胞上观察到大量厚垣孢子,在茎部部分维管束导管中也观察到少量的菌丝,此时48.6%的幼苗表现萎蔫症状;接种后9 d,茎部所有维管束导管都充斥着密集的菌丝体,此时91.7%的幼苗表现枯萎和死亡。与侵染感病品种相比,该菌株在侵染中抗品种和高抗品种时,在侵染时间上要明显滞后。本研究从组织病理学的角度观察并分析了GFP标记的西瓜枯萎病菌侵染西瓜根茎部的过程,为研究西瓜枯萎病菌的致病机理提供参考。     

Anatomy of cranberry stem gall and localization of bacteria in galls

ABSTRACT Cranberry stem gall is characterized by tumors that girdle stems, thereby killing all distal leaves, flowers, and fruit. Bacteria that produce high levels of the plant growth hormone indole-3-acetic acid (IAA) are associated with and believed to cause cranberry stem gall. The anatomy of naturally occurring galls on woody cranberry plants and galls caused by inoculation of micropropagated cranberry plants with Pantoea agglomerans strain 4/99 was consistent with elevated levels of IAA in plants. Field galls exhibited hypertrophy and hyperplasia of tissue external to the vascular cambium, resulting in extensive stem swelling and splitting of the periderm. Similarly, galls on micropropagated plants contained enlarged cortical parenchyma cells. The current year's xylem vessels in field galls were narrow and dense compared with xylem vessels of healthy stems. Curved xylem elements apparently developed de novo within field galls and galls on inoculated plants. Cavities and fissures in both types of galls contained dense aggregates of bacteria. Treatment of micropropagated plants with synthetic IAA caused hypertrophy of cortical parenchyma and formation of adventitious roots. The results support the hypothesis that IAA-producing bacteria cause cranberry stem gall.     

Changes in morphology and measurement of cytokinin levels during the development of witches' brooms on cocoa

The anatomy of diseased stems from brooms induced on cocoa ( Theobroma cacao ) by the fungus Crinipellis perniciosa was examined using the scanning electron microscope, and the amounts of selected cytokinins were measured by immunoassay.
Although the diseased stems were wider in diameter with a lower dry weight:fresh weight ratio than stems of healthy plants, the overall internal organization of tissues remained unchanged, and all the cell types were present. However, the vascular tissue was less differentiated in diseased stems since xylem vessels were absent and the ratio of phloem fibres and sieve tubes to phloem parenchyma was reduced. Cell sizes in diseased stems were larger for all tissues but cell numbers were unchanged except for xylem where fewer cells were seen.
Cytokinins (zeatin, zeatin riboside, isopentenyl adenine and isopentenyl adenosine) were measured in healthy plants and diseased stems at specific stages of disease development. Of these cytokinins only zeatin riboside was present in significantly greater amounts in diseased tissue. The likely role of plant growth regulator balance in stem enlargement and proliferation in witches' broom disease of cocoa is discussed.     

电镜观察镰刀菌在四季豆植株体内侵染过程

用电子显微镜观察镰刀菌(Fusarium oxysporium f.sp.phaseoli)侵染四季豆茎部的过程。结果表明,病菌主要侵染四季豆木质部导管,而同一横切面上的皮层几乎见不到菌体。在同一病株中,基部的导管被病菌严重充塞,越近上部,导管中的病菌越少;同一植株横切面木质部的各导管,病原菌分布不均匀。     

Colonization of Host Plants by the Fire Blight Pathogen Erwinia amylovora Marked with Genes for Bioluminescence and Fluorescence

ABSTRACT To follow the movement of Erwinia amylovora in plant tissue without dissection, this bacterium was marked with either the lux operon from Vibrio fischeri or the gfp gene from the jellyfish Aequorea victoria, both carried on multicopy plasmids and expressed under the control of the lac promoter from Escherichia coli. Movement of the pathogen was visualized in leaves, stems, and roots of apple seedlings, and migration of E. amylovora was traced from inoculation sites in the stem to as far as the roots. Green fluorescent E. amylovora cells were observed in the xylem and later appeared to break out of the vessels into the intercellular spaces of the adjacent parenchyma. Inoculation in the intercostal region of leaves caused a zone of slow necrosis that finally resulted in bacterial invasion of the xylem vessels. Labeled bacteria could also be seen in association with the anchor sites of leaf hairs. Distortion of the epidermis adjacent to leaf hairs created openings that were observed by scanning electron microscopy. As the intercostal region, the bases of leaf hairs provided E. amylovora access to intact xylem vessels, which allowed further distribution of the pathogen in the host plant.     

Fire blight. Why do views on host invasion by Erwinia amylovora differ?

The fire blight pathogen, Erwinia amylovora, commonly infects flowers and shoots of certain rosaceous hosts and systemic (whole‐tree) invasion sometimes follows. The bacterium may be found in the parenchyma of bark tissue and/or in mature xylem vessels of stem tissue. Views differ on initial sites of multiplication and the optimal route for systemic migration. This article presents the evidence on which the different views are based. There are limited observations on orchard pear and apple trees; in most experimental studies, young apple shoots on potted plants were used. Tissue maturity at the site of shoot inoculation is of prime importance. If xylem vessels are damaged, inoculum may be sucked into the vessels and the bacteria will multiply there. In younger tissue, there is less suction pressure. The critical stem entry site for the invasion of cortical parenchyma seems to be near or above the most recently unfolded leaf. No one has suggested that migration in bark tissue cannot be a major route. If the xylem route is followed, the pathogen needs a means of escape into bark tissue, where typical symptoms develop; means of escape from mature xylem vessels have not been demonstrated and remain a matter for speculation. Published evidence does not seem to support the idea that fire blight is a vascular wilt disease, nor that the extracellular polysaccharide produced by E. amylovora is a toxin and responsible for the wilting symptom seen in the early stages of the disease.     

大白菜软腐病潜伏侵染的组织病理学研究

厌气诱发24～48小时后,大白菜叶柄上可以出现由潜伏侵染的软腐欧氏杆菌(Erwinia carotovora subsp.carotovora)引起的水渍状浸离斑。对病斑的邻近组织(病斑以上1～5毫米)断面进行电镜扫描观察,结果指出,病原细菌最初是潜伏在维管束的导管中的。这些细菌生长繁殖后可以消解螺纹导管间的薄壁部分,最后使螺纹松驰解体,再进一步扩展到邻近的薄壁组织为害,表现症状。叶柄表面没有任何自然孔口,但有自然形成的不规则细微裂痕。细菌在完好的叶柄表面不进行分裂繁殖,但在裂痕附近有繁殖和堆积现象。细菌在叶片上的气孔附近没有明显的聚集和繁殖迹象。有菌的气孔会由于被刺激产生的胶状分泌物而阻塞。     

水稻白叶枯病成株抗性品种病理解剖研究

 成株抗性品种(简称成抗品种)苗期感病阶段和感病品种接种后,病菌在叶片内扩展和繁殖速度快,叶片内部组织变态,导管内菌量多、菌体形态正常;成抗品种成株抗病阶段和全生育期抗病品种(简称全抗品种)与此相反,病菌在接种叶片内扩展和繁殖速度慢,叶片内部组织结构正常,导管内菌量较少.并出现了不同形态的物质,将病菌包围,菌体形状不规则,这些物质在健康叶片、成抗苗期接种叶片和感病品种病叶的导管中未观察到,其形成似乎与品种抗性表现有关系.     

2009年造成湖南省水稻大面积矮缩的是南方水稻黑条矮缩病

2009年在湖南省大面积发生一种水稻病毒病,其症状表现为植株矮缩、叶色深绿、高位分蘖、茎秆出现乳白色或浅褐色点条状突起、茎节上出现气生须根。电镜结果表明:在病株韧皮部可见具斐济病毒特征性晶格状排列球状病毒粒体。病毒基因组S10片段部分序列的相似性比对与系统进化树分析表明,该病害是南方水稻黑条矮缩病。     

Phloem and xylem modifications of Vitis vinifera stems in response to flavescence dorée phytoplasma infection

Anatomical modifications of xylem and phloem tissues of grapevine (Vitis vinifera) stems of shoots infected by the flavescence dorée phytoplasma (FDp) were first observed and described in the 1960s, but never quantified in detail. In this paper, we describe and quantify the impact of FDp on grapevine stem tissues, and relate it to the level of expression of symptoms and to cultivar-specific FDp susceptibility. For this purpose, we measured and quantified the anatomical parameters of xylem and phloem tissues of a tolerant (Merlot) and a susceptible (Chardonnay) cultivar. For each cultivar, thin sections of eight shoots with symptoms from FDp-infected grapevines, eight symptomless shoots from the same FDp-infected grapevines, and eight symptomless shoots from symptomless grapevines (control) were compared. Results showed general inhibition of xylem growth and proliferation of phloem tissues (hyperplasia) with lack or irregular arrangement of the fibre-sclereids in the axial phloem of the stems from shoots with symptoms, irrespective of the cultivar. Xylem vessels of infected Merlot shoots were partly occluded by tyloses and a higher number of smaller vessels were produced than in control plants. Thus, the anatomical responses confirmed the detrimental effect of FDp on stems of infected grapevine shoots, including impaired stem development and lack of periderm formation. Statistically significant differences were found between the two cultivars with different levels of susceptibility to FDp infection.     

荧光假单胞菌PEF-5#18防控番茄枯萎病的定殖机理

通过电转法成功将EGFP表达质粒p EGFP20转入对番茄枯萎病具有优良防病效果的荧光假单胞菌PEF-5#18体内,并研究了该生防菌在番茄根际土壤与植株根、茎内部组织的定殖情况。结果表明,处理25 d后,标记型与野生型生防菌处理均能有效控制番茄枯萎病害并增加植株鲜、干重量。与病原菌阳性对照相比,野生型菌株的防治效果为76.92%,鲜重增加194.10%,干重增加564.71%;接种后的PEF-5#18能良好定殖于番茄根际土壤并扩展进入植物根茎内部生长,其分布数量呈现出根际土壤(1.05×106 CFU/g)根(6.75×104 CFU/g)茎(1.25×102 CFU/g);与病原菌阳性对照相比,接种PEF-5#18对番茄根际土壤的可培养细菌总数影响差异不显著,但可显著提高根(增幅为8.42%)、茎(增幅为14.78%)内的可培养细菌总数以及根际土壤pH(增幅为1.61%);接种PEF-5#18能显著降低根际土壤与根、茎内病原菌侵染数量;激光共聚焦镜检发现,接种25 d后,PEF-5#18能大量分布于番茄根系表面、根内木质部、根内皮层细胞、根内细胞间隙、茎内维管及其周围细胞。