荔枝霜疫霉的研究

对华南的荔枝霜疫霉(Peronophythora litchii Chen ex Ko et al)的形态和营养特性进行了研究,并和新模式种进行了比较。发现此菌孢囊梗的生长是一种有限-无限生长类型,或称之为多级有限生长。即孢囊梗上的小分枝大多数是有限生长的,在其顶端同时形成孢子囊。但有时在同一孢囊梗上有的小分枝会继续生长,形成二级、三级甚至四级孢囊梗。在营养要求上与疫霉无大差别,能在天然和合成培养基上旺盛生长,需要硫胺素,ca~(++)和有机二元酸,能利用NH_4~+ 或No_3~-为其氮源,并能利用淀粉为其碳源,菌体匀浆中测出淀粉酶活性。根据孢囊梗的独特生长方式,我们认为完全有理由承认这菌是一个新属,并可成为新科,霜疫霉科。本文中已将霜疫霉科作了修改描述。孢囊梗已被修改为:孢囊梗多级有限生长。无疑这菌是腐霉科和霜霉科的中间类型。在营养类型与有性器官上和疫霉相近,而其孢子囊的形成和霜霉相似。但其孢囊梗的多级有限生长方式则和这两科都不相同。     

开口箭甾体皂甙元的分离鉴定及其抗荔枝霜疫霉菌活性

在以荔枝霜疫霉菌为指示菌的生物活性测定导向跟踪下,运用TLC、硅胶柱层析、反相硅胶柱层析和凝胶柱层析等分离纯化方法,从开口箭(Tupistra chinensis)甲醇提取物的乙酸乙酯分部萃取物中分离纯化得到2个抗菌化合物。经现代光谱(MS、IR、1D NMR和2D NMR)分析,确定化合物1为1β,2β,3β,4β,5β,7α-六羟基螺甾-25(27)-烯-6-酮,化合物2为螺甾-25(27)-烯-1β,2β,3β,4β,5β,6β,7α-七醇。离体抗菌活性测定化合物1和化合物2抑制荔枝霜疫霉菌(Peronophythora litchii)孢子囊萌发的EC50分别为100.28 mg.mL-1和124.37 mg.mL-1。用质量浓度为500 mg.mL-1的两个化合物分别处理后接种霜疫霉菌,供试荔枝果实的发病率分别为16.7%和18.9%。     

苦槛蓝叶的黄酮类成分及其荔枝霜疫霉抑制活性研究

为了解苦槛蓝(Myoporum bontioides)的化学成分,采用色谱分离法从叶中分离得到11个化合物,分别鉴定为：5,7,3’-三羟基-4’-甲氧基黄酮(1)、3,5,7,4’-四羟基-3’-甲氧基黄酮(2)、5,7,4’-三羟基-3’,5’-二甲氧基黄酮(3)、木犀草素(4)、山奈酚(5)、鼠李黄素(6)、5,7-二羟基二氢黄酮(7)、7,4’-二羟基二氢黄酮(8)、5,7,3’,4’-四羟基二氢黄酮(9)、5-O-乙酰基-3,7,3’,4’-四羟基二氢黄酮(10)和7-甲氧基香橙素(11)。除化合物4、7、11之外,其他化合物均为首次从苦槛蓝叶中分离得到。菌丝生长速率法测试表明化合物4、7～9和11对荔枝霜疫霉菌具有较好的抑菌活性。     

CRISPR/Cas9介导的荔枝霜疫霉基因组编辑系统的建立

参考大豆疫霉的CRISPR/Cas9基因组编辑技术体系,针对荔枝霜疫霉RXLR效应蛋白编码基因Pl Avh133设计了20 bp的sgRNA靶向序列,结合同源替换的方式对该基因进行敲除。利用聚乙二醇(PEG)介导的原生质体转化,共获得了58个具有G418抗性的转化子,通过PCR和测序分析证明其中5个转化子的Pl Avh133基因被敲除,敲除效率约为8.6%。荧光定量PCR分析证实Pl Avh133敲除突变体中该基因不表达。本研究结果为荔枝霜疫霉的基因功能研究提供了重要的技术基础。     

绿僵菌SC0924酚酸类代谢产物及其抗荔枝霜疫霉活性

从绿僵菌SC0924固体发酵物中分离得到8个酚酸类化合物,通过波谱分析,分别鉴定为香草酸(1)、丁香酸(2)、邻氨基苯甲酸(3)、苯乙酸(4)、阿魏酸(5)、二氢阿魏酸(6)、2-羟基-3-苯丙酸(7)和2-羟基-3-对羟基苯丙酸丁酯(8).以滤纸片琼脂扩散法对以上化合物进行抗荔枝霜疫霉活性试验,结果表明除化合物2和6外,其余化合物均有抑菌活性.     

绿僵菌SC0924含氮杂环类代谢产物及其抗荔枝霜疫霉活性

从绿僵菌(Metarhizium sp.SC0924)固体发酵物中分离得到12个含氮杂环类化合物,通过波谱分析,分别鉴定为环(丙氨酸-脯氨酸)(1)、环(脯氨酸-酪氨酸)(2)、环(缬氨酸-亮氨酸)(3)、环(亮氨酸-异亮氨酸)(4)、腺嘌呤(5)、腺嘌呤核苷(6)、尿嘧啶(7)、胸腺嘧啶(8)、胸腺嘧啶脱氧核苷(9)、4-乙酰氨基咪唑(10)、焦谷氨酸正丁酯(11)和光敏素(12).滤纸片琼脂扩散法试验表明化合物2和3具有较弱的抗荔枝霜疫霉活性.     

内吸杀菌剂甲霜灵对向日葵霜霉菌发育的影响

利用电镜技术研究了内吸杀菌剂甲霜对向日葵霜霉菌在寄主向日葵上发育的影响,观察发现甲霜引起病菌发生一系列的变化,病菌胞间菌丝细胞内液泡,电子致密体增加,线粒体肿胀畸形,菌丝细胞壁不规则地加厚,部分极度加厚的细胞壁构成了假膜,并且在菌丝上还有不规则的突起形成,病菌吸器外间质明显不规则地变宽,其中充满大量电子致密度高的物质;部分吸器体不能正常扩张膨大,发育受阻而成畸形,向日葵霜霉菌的以上变化导致菌丝细胞     

大白菜感染霜霉菌后细胞超微结构变化的研究

利和透射电子显微镜研究了大白菜抗病品种和感病品种受霜霉菌侵染后叶肉细胞超微结构的变化及其与抗病性的关系。大白菜受霜霉菌侵染后,在抗病品种叶片中,靠近侵染点处的几个细胞由于纤丝化而迅速死亡,此即过敏性死细胞。过敏性细胞坏死是一种主要的抗性机制。在过敏性坏死细胞周围的一些细胞的细胞质中出现大量吞噬体和质膜体;线粒体膨大成长条形,其内嵴的数量增多,高尔基体小泡也大量增加;核糖体丰富;内质网膨大。这些超微     

Fungicide Effectiveness during the Various Developmental Stages of Peronophythora litchii In Vitro

Litchi downy blight caused by Peronophythora litchii is one of the most destructive diseases suffered by litchi in China. This study has evaluated the activities of the fungicides dimethomorph (DMM), azoxystrobin (AZB), famoxadone (FMD), metalaxyl (MTL), cymoxanil (CYX) and mancozeb (MCB) on the mycelial growth, sporulation, zoospores release, and germination of sporangia as well as of cystospores of P. litchii . Dimethomorph and MTL inhibited mycelial growth more effectively than the other fungicides tested. Mycelial growth was affected less by MCB, and only weakly by AZB, FMD and CYX. Sporangia production was more strongly reduced by DMM and MTL than by AZB, FMD, CYX or MCB. Zoospore release from the sporangia was most sensitive to AZB and FMD, less sensitive to MCB and insensitive to DMM, MTL and CYX. Direct germination of sporangia of P. litchii was the most strongly inhibited by AZB and FMD, followed by DMM and MCB, whilst the sensitivity of this life stage to MTL and CYX was relatively low. Germination of encysted zoospores of P. litchii was the most sensitive to AZB and FMD, followed by DMM. Mancozeb was moderately active, while MTL and CYX did not inhibit this development stage. This is the first report on the in vitro response of the litchi pathogen, P. litchii, to fungicides. These findings can be valuable tools in setting up efficient disease management programmes to control litchi downy blight.     

荔枝若干重要果实品质性状的QTL分析

以‘马贵荔＇ב焦核三月红＇76株F_1代群体为试材,检测了酒石酸、苹果酸、蔗糖含量和单果重4个果实性状分离的情况。结果表明,4个性状表现为连续分布,具有数量性状的典型特征,与4个果实性状连锁的QTL位点23个,其中控制酒石酸的QTL为2个,控制苹果酸的QTL为4个,控制蔗糖的QTL为12个和控制单果重的QTL为5个。各QTL的LOD值在3.15～5.61之间,可解释13.85%～88.3%的表型变异。     

LcEIL2/3 are involved in fruitlet abscission via activating genes related to ethylene biosynthesis and cell wall remodeling in litchi

Fruit crops are subject to precocious fruit abscission, during which the phytohormone ethylene (ET) acts as a major positive regulator. However, the molecular basis of ET‐induced fruit abscission remains poorly understood. Here, we show that two ETHYLENE INSENSITIVE 3‐like (EIL) homologs in litchi, LcEIL2 and LcEIL3, play a role in ET‐activated fruitlet abscission. LcEIL2/3 were significantly upregulated in the fruit abscission zone (AZ) during the ET‐induced fruitlet abscission in litchi. The presence of LcEIL2/3 in wild‐type Arabidopsis and ein3 eil1 mutants can accelerate the floral organ abscission. Moreover, the electrophoretic mobility shift assay and dual luciferase reporter analysis illustrated that LcEIL2/3 directly interacted with the gene promoters to activate the expression of cell wall remodeling genes LcCEL2/8 and LcPG1/2, and ET biosynthetic genes LcACS1/4/7 and LcACO2/3. Furthermore, we showed that LcPG1/2 were expressed in the floral abscission zone of Arabidopsis, and constitutive expression of LcPG2 in Arabidopsis promoted the floral organ abscission. In conclusion, we propose that LcEIL2/3 are involved in ET‐induced fruitlet abscission via controlling expression of genes related to ET biosynthesis and cell wall remodeling in litchi.     

Multiple shoot induction and plant regeneration in litchi (Litchi chinensis Sonn.)

Multiple shoot induction in Litchi chinensis Sonn. (litchi) has been achieved by two methods: (1) direct germination of litchi seeds in 6-benzylaminopurine (20 mg l^–1)-supplemented MS liquid medium and supported on a filter-paper bridge and (2) in planta treatment with 6-benzylaminopurine (100 μg on alternate days) of the axillary bud regions of plants germinated and maintained under sterile conditions. While the former method resulted in as many as 27.5±8.6 shoot buds from the cotyledonary node within 4 weeks, the latter yielded on average approximately 8 shoot buds from each treated node in 8 weeks. The cytokinin treatment in planta consisted of placing sterile filter paper moistened with sterile distilled water over the node and adding different concentrations of 6-benzylaminopurine. Both methods of multiple shoot induction were found to be effective for the five genotypes of litchi that were tested. The shoots elongated and rooted directly in vermiculite after a pulse treatment with IBA (25 mg/ml) for 15 min. Fungus growth which is a serious problem in litchi tissue culture, was controlled using a fungicide, Bavistin, and by eliminating organic nutrients from the growth medium. Received: 27 July 1998 / Revision received: 30 September 1998 / Accepted: 27 October 1998     

低温对荔枝果肉膜脂过氧化和保护酶活性的影响

对‘白蜡’荔枝果肉在3℃和-1℃下的贮藏效果及果肉膜脂过氧化和保护酶活性进行了研究。结果表明,去果皮的荔枝果肉在3℃下贮藏40d,-1℃下保鲜50d,不表现冷害症状。低温下果肉的细胞膜透性、丙二醛(MDA)含量逐渐增加。冷藏30d,-1℃与3℃果肉之间的膜透性、MDA含量没有显著差异,但30d后,-1℃处理明显延缓了果肉膜透性和MDA含量的增加。在3℃下贮藏果肉的超氧化物歧化酶(SOD)和抗坏血酸过氧化物酶(ASA-POD)和过氧化氢酶(CAT)活性在20d、过氧化物酶(POD)活性在30d时分别到达高峰值。-1℃延缓了这些保护酶活性的升高并降低了峰值。     

采后荔枝果实冷害过程中多胺含量的变化

以桂味荔枝果实为材料,研究冷害及外源亚精胺(Spd)处理对果实膜透性和内源多胺含量变化的影响。结果表明,在0℃下贮藏时果实发生冷害过程中,荔枝果皮中腐胺(Put)、Spd、精胺(Spm)含量在14d后明显增加,膜透性快速增大,21d时果皮出现明显的冷害褐变,Put进一步积累,而Spm含量下降,Spd保持较高水平。非冷害温度(3℃)下贮藏时,果皮多胺含量变化相对较小。0℃下果肉的多胺含量和变化幅度低于3℃果实,并延迟7d衰老。外源Spd处理明显提高果实内源多胺含量的同时,延缓了果皮相对膜透性增加,减轻了冷害。这表明果皮中Put的积累可能是荔枝果实冷害的结果,冷藏初期Spm含量的上升可能是果实对冷害的防卫反应。     

荔枝树干液流速率与气象因子的关系

采用热扩散茎流仪于2011—2012年连续监测‘桂味’荔枝树干液流速率,将所得数据和果园内自动气象站观测数据一起进行对比分析,建立了液流速率与气象因子的关系模型。实验结果表明:(1)树干液流速率日变化呈现"昼高夜低"的规律。季节变化有呈现"先高后低"的趋势,整体上以果实发育期和成熟期(5—7月)液流速率较高。2011年年平均液流速率明显大于2012年;(2)树干液流速率日变化在晴天时多呈单峰曲线,且振幅较大;在雨天呈多峰曲线,且振幅较小。总体上,晴天平均液流速率约是雨天的两倍;(3)回归分析表明,在一定范围内,树干液流速率与太阳辐射强度、空气温度呈正相关,与空气湿度呈负相关。影响液流速率最重要的气象因子在晴天是空气温度,在雨天是太阳辐射。     

大核和焦核"桂味"荔枝果实发育及其发育期间果皮中内源激素含量的变化比较

同一株大核和焦核"桂味"荔枝的果实生长型均呈"单S型",其果实大小差异主要由种子大小不同引起.果实发育期间,两者果皮中内源激素变化的规律大体上一致,大核果皮中GA3、IAA含量和(IAA ZRs GA3)/ABA比值均高于焦核果皮,但ZRs含量和ZRs/ABA比值则比其低,ABA含量的差异无规律可循.     

裂果易发性不同的荔枝品种果皮中细胞壁代谢酶活性的比较

“糯米糍”荔枝裂果率极显著高于“淮枝”,前者果皮中的果胶酶、纤维素酶和果胶甲酯酶的活性高于后者,其中以果胶酶活性差异最明显,其次是纤维素酶,果胶甲酯酶差异最小;“糯米糍”细胞壁结合型的过氧化物酶(POD)和多酚氧化酶(PP0)活性明显高于“淮枝”,而水溶性POD和PP0的活性则两个品种间无明显差异。据此认为,果皮细胞壁水解酶活性以及细胞壁结合型的POD和PPO的活性高的荔枝品种,其裂果率也高。文章对细胞壁代谢相关酶类在果皮抗裂性形成中的作用进行了讨论。