芒果炭疽病菌环境pH信号调控基因PalF的克隆与分析

通过本地查找胶孢炭疽病菌(Colletotrichum gloeosporioides)橡胶菌株全基因组测序数据,获得PalF基因的序列信息。利用同源克隆的方法扩增胶孢炭疽病菌芒果菌株PalF基因,获得片段大小为2 369 bp的目的基因片段。对该基因序列进行分析发现,该片段含有完整的开放式阅读框,与橡胶炭疽菌PalF基因的序列同源性达100%。用RT-PCR的方法获得PalF基因的cDNA序列,序列进行分析发现该基因全长为2 310 bp,其中含有1个大小为59 bp的内含子,推测编码769个氨基酸。对PalF基因的氨基酸序列进行推测保守结构域分析可知PalF蛋白中含有Arrestin-C保守结构域,推测该基因参与环境pH感应信号转导途径(Ambient pH-sensing signal transduetion pathway)的调控,为进一步研究该基因在环境pH感应信号转导途径中的作用奠定了基础。     

墨兰品种健叶与感染胶孢炭疽菌病叶的表面结构及氨基酸含量比较

【目的】明确墨兰品种健叶与感染胶孢炭疽菌病叶的叶片表面结构及氨基酸含量的变化,为墨兰品种抗病性利用提供理论依据。【方法】以中感墨兰品种太平洋为样本,利用扫描电镜观察感染胶孢炭疽菌墨兰叶片表面超微结构的变化;以不同感病水平墨兰品种金华山(低感品种)、太平洋(中感品种)和万代福(高感品种)为样本,利用氨基酸自动分析仪检测感病墨兰叶片除色氨酸外其余17种氨基酸含量的变化。【结果】中感墨兰品种太平洋感染胶孢炭疽菌后,叶片表面超微结构发生明显改变,蜡质层破裂,表皮细胞受损,气孔功能渐失,严重影响叶片功能与整体观感。3种不同感病水平的墨兰品种感染胶孢炭疽菌后,氨基酸种类未发生改变,但总氨基酸含量均明显提高,其中以低感墨兰品种金华山提高最多,高感墨兰品种万代福提高最少,3个品种间差异显著;各种氨基酸在不同感病水平墨兰品种间的变化量差异显著,甲硫氨酸(Met)在高感墨兰品种中变化量最大,胱氨酸(Cys)在中感墨兰品种中变化量最大,天门冬氨酸(Asp)、组氨酸(His)和赖氨酸(Lys)在高感与低感墨兰品种中变化量较大,甘氨酸(Gly)和苏氨酸(Thr)在中感与低感墨兰品种中变化量较大,其他10种氨基酸则均在低感墨兰品种中变化量最大;丙氨酸(Ala)、谷氨酸(Glu)、苯丙氨酸(Phe)、缬草氨酸(Val)、丝氨酸(Ser)、精氨酸(Arg)和苏氨酸(Thr)7种氨基酸是影响墨兰品种感病强弱的主要氨基酸种类。【结论】胶孢炭疽菌侵染影响墨兰品种叶片表面结构和氨基酸含量,营养物质变化量与墨兰品种感病水平高低有关。     

Development of the infection strategy of the hemibiotrophic plant pathogen,Colletotrichum orbiculare,and plant immunity

The hemibiotrophic fungus Colletotrichum orbiculare forms appressoria as infection structures and primarily establishes biotrophic infection in cucumber epidermal cells. Subsequently, it develops necrotrophic infection. In the pre-invasion stage, morphogenesis of appressoria of C. orbiculare is triggered by signals from the plant surface. We found that C. orbiculare PAG1 (Perish-in-the-Absence-of-GYP1), a component of MOR [morphogenesis-related NDR (nuclear Dbf2-related) kinase network] plays an essential role as a key component of the plant-specific signaling pathway and that hydrolysis of cutin by a spore surface esterase creates a cutin monomer that constitutes a key plant-derived signal. Development of the infection structure of C. orbiculare is strictly regulated by the cell cycle and we found that proper regulation of G1/S progression via two-component GAP genes, consisting of BUB2 (Budding-Uninhibited-by-Benomyl-2) and BFA1 (Byr-Four-Alike-1) is essential for the establishment of successful infection. In the post-invasion stage, the establishment of the biotrophic phase of hemibiotrophic fungi is crucial for successful infection. We found that C. orbiculare WHI2 (WHIsky-2), an Saccharomyces cerevisiae stress regulator homolog, is involved in the phase transition from biotrophy to necrotrophy through TOR (Target of Rapamycin) signaling, and is thus essential for full pathogenesis.     

热处理对2种潜伏炭疽菌生长和致病性的影响

 通过测定热处理对潜伏侵染于芒果果实中的胶胞炭疽菌(Colletotrichum gloeosporioides Penz.)和香蕉果实中的芭蕉炭疽菌[C.musae(Berk.& Curt.)Arx.]离体培养菌的生长、繁殖和致病性的影响,结果表明:当温度达55℃和60℃,时间20 min时,对菌体的生长和孢子萌发可起明显抑制或杀伤作用,并降低其致病性。2种炭疽菌中,芭蕉炭疽菌比胶胞炭疽菌对热更敏感。作者认为,果实采后热处理时,应根据果实种类和不同菌,采用不同的处理温度和时间,才能得到更好的效果。     

Breeding for resistance to coffee berry disease in Coffea arabica L. II. Inheritance of the resistance

Summary The inheritance of resistance to coffee berry disease (CBD) has been studied by applying a preselection test to F₂ progenies of a half diallel cross between 11 coffee varieties with different degrees of resistance and to sets of parental, F₁, F₂, B₁₁ and B₁₂ generations of crosses between resistant and susceptible varieties. True resistance to CBD appears to be controlled by major genes on three different loci. The highly resistant variety Rume Sudan carries the dominant R- and the recessive K-genes. The non-allelic interaction between these two genes is of a duplicate nature. The R-locus has multiple alleles with R ₁R₁alleles present in Rume Sudan and the somewhat less effective R ₂R₂alleles in a variety like Pretoria, which also has the K-gene. The moderately resistant variety K7 carries only the recessive K-gene. The arabica-like variety Hibrido de Timor (a natural interspecific arabica x robusta hybrid) carries one gene for CBD resistance on the T-locus with intermediate gene action. It probably inherited this gene from its robusta parent. There is circumstantial evidence that the resistance to CBD is of a stable nature, but it is advisable to accumulate in one genotype as many resistance genes as possible by combining in the breeding programme the resistance of Rume Sudan with that of Hibrido de Timor.     

AFLP analysis of genetic variability among Stylosanthes guianensis accessions resistant and susceptible to the stylo anthracnose

Stylosanthes guianensis, belonging to the genus Stylosanthes, is one of the most important tropical forage legumes and is native to South and Central America and Africa. Anthracnose, caused by the fungus Colletotrichum gloeosporioides (Penz.) Sacc., is a major constraint to the extensive use of Stylosanthes as tropical forage. Forty‐two accessions of S. guianensis were assessed with amplified fragment length polymorphism (AFLP) for genetic diversity and for resistance to anthracnose. In AFLP analysis, four selective primer combinations screened from 96 primer combinations were used to analyse these accessions, and a total of 225 clear bands were used for genetic similarity (GS) analysis, showing a 95.5% level of polymorphism on average. GS from 31.0% to 95.0% among the accessions was calculated with ntsys ‐pc software. The dendrogram was constructed with unweighted pair group method of averages (UPGMA) based on the AFLP data, and five clusters were defined at 48% GS. Two typical strains of C. gloeosporioides from Stylosanthes in China were used for anthracnose resistance screening. Most of the plant accessions showed variation in the reaction to two strains, and the correlation of resistance had a value of 0.904 (P < 0.01), suggesting common resistance to the two strains. The resistance accessions were randomly distributed in different groups of UPGMA clustering. These results demonstrate that AFLP analysis is an efficient method for evaluating the genetic diversity among S. guianensis accessions.     

Use of molecular markers to accelerate the breeding of common bean lines resistant to rust and anthracnose

The main goal of this work was to introduce resistance genes for rust, caused by Uromyces appendiculatus, and anthracnose, caused by Colletotrichum lindemuthianum, in an adapted common bean cultivar through marker-assisted backcrossing. DNA fingerprinting was used to select plants genetically closer to the recurrent parent which were also resistant to rust and to race 89 of C. lindemuthianum. DNA samples extracted from the resistant parent (cv. Ouro Negro), the recurrent parent (cv. Rudá), and from BC₁, BC₂ and BC₃ resistant plants were amplified by the RAPD technique. The relative genetic distances in relation to the recurrent parent varied between 9 and 59% for BC₁, 7 and 33% for BC₂, and 0 and 7% for BC₃ resistant plants. After only three backcrosses, five lines resistant to rust and anthracnose with, approximately, 0% genetic distance in relation to the recurrent parent were obtained. These lines underwent field yield tests in two consecutive growing seasons and three of them presented a good yield performance, surpassing in that sense their parents and most of the reference cultivars tested.     

田七炭疽病的病原及其生物学特性的研究

炭疽病是靖西县田七〔Panax notoginseng(Burk.)F.H.Chen〕的主要病害,其病原有以C_1、C_2、C_3和C_4菌株为代表的四种形态型。经鉴定,C_1和C_2菌株为Colletotrichum gloeosporioides Penz.,C_3和C_4为C.dematium(Pers.ex Fr.)Grove。过去认为靖西田七幼苗猝倒病是由丝核菌Rhizoctonia sp.引起的,经研究证实,其病原是C.gloeosporioides引起的一种幼苗炭疽病。 C_1菌株是田七炭疽病的优势菌株,其生长温度范围4～32℃,最适温度为15～28℃;水滴或水膜是分生孢子形成和萌发的主要条件。C_1菌株在PDA培养基上有产孢型与不产孢型之分,但用“植物组织+PDA”培养不产孢型,可在植物组织上诱发产孢成功,其中以茄科和豆科的一些植物组织上产孢量较大。     

草莓炭疽病病原鉴定及其生物学特性研究

于辽宁省抚顺市栽培的草莓品种的茎上分离得到一种炭疽病菌。针对病菌的形态特征、培养性状及其有性形态进行了生物学特性等研究。结果表明:该炭疽病菌属胶孢炭疽菌〔Colletotrichum gloeosporioides(Penz.)Sacc.〕;该病原菌在PSA和PDA培养基上生长最好,PSA培养基最适于病菌产孢;菌丝生长和产孢的温度范围为10～35℃,最适温度25℃,适宜pH值为4～6;光照可促进产孢,而全黑暗有抑制作用;对碳源的利用,果糖最好,纤维素最差;对氮源的利用,蛋白胨最好,尿素最差;孢子萌发的最适温度为30℃,低于5℃不能萌发;萌发最适pH值为5～7,高于10不萌发,光照对孢子萌发无明显影响。     

宁夏枸杞炭疽病病原的生物学特性研究

对宁夏枸杞炭疽病原(Colletotrichum gloeosporioides Penz)的生物学特性进行了研究.结果表明:22～31 ℃为最适温度,有利菌落生长、产孢,接种孢子6 h可萌发.相对湿度90%(RH)以上时,有利孢子萌发.碳源以葡萄糖、甘露糖对菌落生长有利,可溶性淀粉对产孢有利.氮源以蛋白胨菌落生长最快,硝酸钾、蛋白胨、尿素产孢量较大.孢子喜在弱酸条件下萌发,pH值4～5时有利菌落生长.连续光照有利菌落生长、产孢.10℃以下,40℃以上不能正常产孢,萌发.致死温度为55 ℃.