几种瓜类作物对根际高温胁迫的生理响应

以黑籽南瓜、‘春秋王2号’黄瓜、‘傲美’苦瓜、‘兴蔬’丝瓜、‘五叶香’丝瓜为试材,采用营养液栽培法,研究了这几种瓜类作物在根际35℃高温处理下的生长量、光合参数及地上部、地下部的渗透胁迫物质的变化。结果显示：处tE5d后,两种丝瓜幼苗的株高、茎粗、叶面积、根长、地上干重及地下干重,叶片相对叶绿素含量、净光合速率（Pn）,气孔导度（Gs）、胞间CO2浓度（Gi）、蒸腾速率（Tr）与对照相比变化均不大,而另外三种瓜类的以上指标均受到明显的抑制;高温使得黑籽南瓜及黄瓜叶片和根系中都积累了大量的膜脂过氧化产物丙二醛（MDA）,渗透调节物质脯氨酸和可溶性蛋白含量在叶片中增大,但在根系中却减小,而处理下‘五叶香’丝瓜的MDA含量与对照差异不显著,且两种渗透调节物质表现为叶片中减小根系中增大;处理下‘兴蔬’丝瓜根系中MDA的减少量及可溶性蛋白的增加量都大于‘傲美’苦瓜。结果表明,根际高温下两个丝瓜品种,特别是‘五叶香’丝瓜有较高的耐受力,通过提高根系中的渗透调节物质含量来降低根际高温对植株整体造成的伤害。     

Physiological basis of different allelopathic reactions of cucumber and figleaf gourd plants to cinnamic acid

To provide an insight into the mechanism of interspecific interactions mediated by allelochemicals, cucumber and figleaf gourd seedlings were compared on their response to cinnamic acid, an autotoxin from root exudates of cucumber. Reactive oxygen species metabolism and plasma membrane H(+)-ATPase activity were examined in roots upon exposure to cinnamic acid. This exposure resulted in significant increases in activities of NADPH oxidase, superoxide dismutase, guaiacol peroxidase, and catalase, as well as in O(2)(.-) production and H(2)O(2) content, in cucumber roots but not in figleaf gourd roots. Notably, the cucumber roots produced significant amount of reactive oxygen species (ROS) immediately after cinnamic acid treatment, consequently increasing membrane peroxidation, decreasing membrane H(+)-ATPase activity, and losing root viability. By contrast, no such changes were observed in figleaf gourd roots. All these results indicated that there was an interspecies difference in the recognition of allelochemicals, which induced oxidative stress accompanied by root cell death in cucumber, an autotoxic plant, but not in figleaf gourd, a cucumber relative.     

Molecular phylogenetic analysis of actin genic regions fromAchlya bisexualis (Oomycota) andCostaria costata (Chromophyta)

Summary Actin genic regions were isolated and characterized from the heterokont-flagellated protists,Achlya bisexualis (Oomycota) andCostaria costata (Chromophyta). Restriction enzyme and cloning experiments suggested that the genes are present in a single copy and sequence determinations revealed the existence of two introns in theC. costata actin genic region. Phylogenetic analyses of actin genic regions using distance matrix and maximum parsimony methods confirmed the close evolutionary relationship ofA. bisexualis andC. costata suggested by ribosomal DNA (rDNA) sequence comparisons and reproductive cell ultrastructure. The higher fungi, green plants, and animals were seen as monophyletic groups; however, a precise order of branching for these assemblages could not be determined. Phylogenetic frameworks inferred from comparisons of rRNAs were used to assess rates of evolution in actin genic regions of diverse eukaryotes. Actin genic regions had nonuniform rates of nucleotide substitution in different lineages. Comparison of rates of actin and rDNA sequence divergence indicated that actin genic regions evolve 2.0 and 5.3 times faster in higher fungi and flowering plants, respectively, than their rDNA sequences. Conversely, animal actins evolve at approximately one-fifth the rate of their rDNA sequences.     

pib基因启动子及其诱导启动性初探

将pib基因上游5.7 kb区段取代pCAMBIA1301中gus基因上游的35S启动子构建了pib拟启动区-GUS+ 35S-hpt 基因表达载体pNAR604。经农杆菌介导转化水稻成熟胚愈伤,获得了转基因抗潮霉素愈伤和36株转基因水稻植株。 转基因抗性愈伤和转基因植株根的组织化学GUS活性检测表明,光照培养下的抗性愈伤和转基因植株根不能使X-gluc显色,而暗处理24 h后的抗性愈伤和定植后转基因植株的根能使X-gluc显色。转基因植株GUS荧光定量分析结果表明,GUS表达具有器官特异性,黑暗处理前根的GUS活性最高、茎次之,分别是是叶片的7倍和3倍,叶片中仅有痕量本底。24 h黑暗处理后根、茎、叶中GUS活性都有增加,且叶片中的增加比例最大,其活性仅次于根。5 mmol/L水杨酸和0.3 mol/L NaCl叶面喷施转基因植株24 h后叶片中GUS活性分别为处理前的2.7和3.6倍。初步确定pib拟启动区是一个诱导型启动子。黑暗、水杨酸和NaCl能诱导该启动子启动活性。     

丝瓜作砧木提高黄瓜耐涝性的研究

研究了涝害胁迫下黄瓜自根苗、南瓜砧嫁接苗和丝瓜砧嫁接苗的耐涝性能。结果表明：涝害胁迫时,丝瓜砧嫁接苗能维持较高的根系活力与较高的叶绿素含量;根系电解质渗出率、丙二醛（MDA）含量和超氧阴离子自由基（O2-）的含量 显著低于黄瓜自根苗和南瓜砧嫁接苗;根系SOD、POD和CAT等细胞保护酶活性高于黄瓜自根苗和南瓜砧嫁接苗;丝瓜作砧木可以提高黄瓜的耐涝性。     

丝瓜作砧木提高黄瓜耐涝性的研究

研究了涝害胁迫下黄瓜自根苗、南瓜砧嫁接苗和丝瓜砧嫁接苗的耐涝性能。结果表明：涝害胁迫时,丝瓜砧嫁接苗能维持较高的根系活力与较高的叶绿素含量;根系电解质渗出率、丙二醛(MDA)含量和超氧阴离子自由基(O2^-)的含量显著低于黄瓜自根苗和南瓜砧嫁接苗;根系SOD、POD和CAT等细胞保护酶活性高于黄瓜自根苗和南瓜砧嫁接苗;丝瓜作砧木可以提高黄瓜的耐涝性。     

亚精胺诱导黄瓜幼苗对白粉病抗性的研究

以黄瓜品种‘长春密刺’幼苗为材料,研究了亚精氨(Spd)诱导黄瓜幼苗对白粉病的抗性,并测定Spd处理和白粉菌接种对黄瓜叶片4种防御酶活性及3种防卫基因表达的影响。结果显示:(1)0.2～1.0mmol.L-1 Spd对黄瓜幼苗白粉病抗性均有不同程度的诱抗效果,并以0.8mmol.L-1 Spd处理效果最明显,诱导效率可达55.3%。(2)喷施Spd或接种白粉菌均可提高黄瓜叶片过氧化物酶(POD)、苯丙氨酸解氨酶(PAL)、几丁质酶和β-1,3-葡聚糖酶的活性,且诱导并接种处理的植株叶片上述酶活性均比只诱导不接种处理的上升速度更快;同时,Spd处理和接种白粉菌可以提高植株叶片中POX、PAL、PR-1a基因的表达量。研究表明,Spd处理可以诱导防卫基因表达的增强,提高防御酶活性,显著降低病情指数,增强黄瓜幼苗对白粉病的抗性。     

Effect of larval density and host species on preimaginal development of Bactrocera tau (Diptera: Tephritidae)

The fruit fly, Bactrocera tau (Walker) (Diptera: Tephritidae), is an important pest of fruit and vegetable crops. In this study, host preference of B. tau females and the effects of host species and larval density on larval survival, pupal weight, adult emergence, and developmental duration were investigated on cucumber (Cucumis sativus L.), sponge gourd (Luffa cylindrical L. (Roem)), bitter gourd [Momordica charantia (Cucurbitaceae) L.], guava [Psidium guajava (Myrtaceae) L.], and tangerine [Citrus reticulata (Rutaceae) (Blanco)]. The results showed that females preferred to cucumber over other host species. Larval feeding experience affected subsequent host oviposition preference of adult females. Host species and initial larval density affected certain aspects of the biology of B. tau. Larval density negatively affected insect performance. Survival rates at low densities were significantly higher than that at high densities. Total developmental duration reduced at high larval densities. Cucumber was more suitable to larval growth. Larvae on cucumber grew faster and the puparia were heavier than that on other host species. Larval survival, pupation rate and adult emergence were higher on cucumber compared with those in other host species. Oviposition preference of adult females was correlated with performance of their offspring.     

Sunflower as a trap crop for the European tarnished plant bug (Lygus rugulipennis)

The European tarnished plant bug, Lygus rugulipennis (Heteroptera: Miridae), inflicts serious damage to glasshouse crops, including cucumber, in which it causes distorted foliage, dead growing points and malformed fruits. In a research involving commercial growers, we tested the possibility of using a trap crop to control L. rugulipennis. We screened the attraction of sunflower and lucerne as trap crops using an olfactometer. Adults (females and males) were more attracted to the odour of either flowering sunflower or lucerne than flowering cucumber. In a glasshouse trial, potted flowering sunflowers were evaluated as a trap crop placed at the ends of each cucumber row. Although the trap crop showed much higher attractiveness than the cucumber crop, this effect was not sufficient to provide an acceptable level of control according to the commercial growers. The growers suggested developing artificial sunflower odour as a more efficient lure for removal trapping. As a first step in this development, we tested the attraction of the headspace odour collected from sunflower in olfactometer trials. Sunflower headspace attracted a higher number of adults compared to a blank or a flowering cucumber. The sunflower odour was analysed by gas chromatography coupled to mass‐spectrometry to determine compounds possibly involved in L. rugulipennis attraction. The chemical analysis of the plant odour showed a well‐defined differentiation between sunflower and cucumber, with a number of monoterpenes released exclusively by sunflower. This, plus an emission rate from sunflower being at least four times more abundant, opens the possibility of using synthetic sunflower volatiles to attract L. rugulipennis within a cucumber background.     

Temperature effects on the reactive oxygen species formation and antioxidant defence in roots of two cucurbit species with contrasting root zone temperature optima

Suboptimal root zone temperature (14°C) was imposed on chilling-sensitive cucumber (Cucumis sativus L.) and chilling-tolerant figleaf gourd (Cucurbita ficifolia Bouché) plants. Exposure of roots to low temperature for up to 10 days caused a strong growth inhibition in cucumber compared with figleaf gourd. Physiological analysis showed that generation of reactive oxygen species (ROS) such as hydrogen peroxide and superoxide anion was significantly induced in cucumber plants as fast as 1 day after low root zone temperature treatment. In addition to the significant induction of antioxidant superoxide dismutase activity, low root zone temperature also increased the mitochondrial electron transport allocated to alternative pathway while decreased cytochrome pathway salicylhydroxamic acid-resistant respiration. However, these defense responses could not compensate for the ROS production, resulting in membrane lipid peroxidation and loss of root cell viability in the low root zone temperature treated cucumber roots. In contrast, 14°C root zone temperature had no significant effects on figleaf gourd plant growth, antioxidant enzymes, ROS levels and alternative respiratory pathway. Hence, difference in ROS metabolism would be associated with the remarkable difference in adaptability of cucumber and figleaf gourd plants in response to suboptimal root zone temperature condition.     

Na2CO3处理对黄瓜嫁接苗与自根苗耐盐性的影响

通过Na2CO3,胁迫对黄瓜嫁接苗和自根苗耐盐性差异的研究,揭示黄瓜幼苗的耐盐机制,为设施黄瓜生产提供理论依据。实验以孝感早瓠瓜为砧木,津春四号黄瓜为接穗,采用不同浓度Na2CO3,溶液对黄瓜嫁接苗和自根苗进行处理,研究其对黄瓜幼苗的生理胁迫效应。结果表明：在0—7000mg／L范围内,随着Na2CO3,处理浓度的增加,黄瓜嫁接苗和自根苗的株高、茎粗、叶面积、地上部鲜重、根鲜重、叶绿素和根系活力均下降,但嫁接苗受抑制的程度显著低于自根苗;脯氨酸、丙二醛和根冠比均呈上升趋势,且这三项指标均表现为嫁接苗显著高于自根苗;叶片SOD酶活性均呈先上升后下降趋势,均在Na2CO3,处理浓度为1000mg／L时达到最大值,且嫁接苗的活性显著高于自根苗。嫁接苗的耐盐性优于自根苗,Na2CO3胁迫条件下,嫁接苗和自根苗的生长条件盐浓度以不超过3000mg／L为宜。     

Isolation and characterization of cDNA clones encoding ADP-glucose pyrophosphorylase (AGPase) large and small subunits from chickpea (Cicer arietinum L.)

Four cDNA clones encoding two large subunits and two small subunits of the starch regulatory enzyme ADP-glucose pyrophosphorylase (AGPase) were isolated from a chickpea (Cicer arietinum L.) stem cDNA library. DNA sequence and Southern blot analyses of these clones, designated CagpL1, CagpL2 (large subunits) and CagpS1 and CagpS2 (small subunits), revealed that these isoforms represented different AGPase large and small subunits. RNA expression analysis indicated that CagpL1 was expressed strongly in leaves with reduced expression in the stem. No detectable expression was observed in seeds and roots. CagpL2 was expressed moderately in seeds followed by weak expression in leaves, stems and roots. Similar analysis showed that CagpS1 and CagpS2 displayed a spatial expression pattern similar to that observed for CagpL2 with the exception that CagpS1 showed a much higher expression in seeds than CagpS2. The spatial expression patterns of these different AGPase subunit sequences indicate that different AGPase isoforms are used to control starch biosynthesis in different organs during chickpea development.     

链格孢Alternaria alternata侵染导致瓜类幼苗白化的机制

用分离自无症状瓜类植物的27株链格孢属Alternaria真菌接种黄瓜,有14株引起了黄瓜幼苗的白化,这些菌株均属于链格孢A. alternata。为揭示A. alternata引起黄瓜幼苗白化的机制,对代表菌株5F29经固体发酵后提取其次级代谢产物,将粗提物按一定量混入基质进行盆栽实验。结果表明,黄瓜、丝瓜和甜瓜对该粗提物敏感,出现白化苗;小白菜和辣椒对粗提物不敏感,生长正常。植物幼苗用粗提物处理的结果与用产生菌接种的结果相一致。粗提物经硅胶柱层析、葡聚糖凝胶柱层析及高效液相色谱（HPLC）等方法分离纯化得到一种活性化合物,经测定该化合物引起黄瓜幼苗白化的最低浓度为12.5μmol/L。该化合物在此浓度下可引起黄瓜幼苗生长点白化,在15μmol/L的浓度下可引起黄瓜幼苗大部分组织白化,致使幼苗很快死亡。应用核磁共振和质谱技术,确定了该化合物的结构,鉴定为一种植物毒素——腾毒素（tentoxin）。研究结果揭示了A. alternata引起瓜类幼苗白化的机制以及不同植物对于腾毒素的不同敏感性。     

Isolation and Characterization of the Genes Encoding Basic and Acidic Chitinase in Arabidopsis thaliana

Plants synthesize a number of antimicrobial proteins in response to pathogen invasion and environmental stresses. These proteins include two classes of chitinases that have either basic or acidic isoelectric points and that are capable of degrading fungal cell wall chitin. We have cloned and determined the nucleotide sequence of the genes encoding the acidic and basic chitinases from Arabidopsis thaliana (L.) Heynh. Columbia wild type. Both chitinases are encoded by single copy genes that contain introns, a novel feature in chitinase genes. The basic chitinase has 73% amino acid sequence similarity to the basic chitinase from tobacco, and the acidic chitinase has 60% amino acid sequence similarity to the acidic chitinase from cucumber. Expression of the basic chitinase is organ-specific and age-dependent in Arabidopsis. A high constitutive level of expression was observed in roots with lower levels in leaves and flowering shoots. Exposure of plants to ethylene induced high levels of systemic expression of basic chitinase with expression increasing with plant age. Constitutive expression of basic chitinase was observed in roots of the ethylene insensitive mutant (etr) of Arabidopsis, demonstrating that root-specific expression is ethylene independent. Expression of the acidic chitinase gene was not observed in normal, untreated Arabidopsis plants or in plants treated with ethylene or salicylate. However, a transient expression assay indicated that the acidic chitinase promoter is active in Arabidopsis leaf tissue.     

低温胁迫对以野生黄瓜(棘瓜)为砧木的黄瓜嫁接苗生理生化指标的影响

在5℃低温胁迫条件下,对以野生黄瓜——棘瓜(Sicyos angulatus L．)和以黑籽南瓜为砧木的黄瓜嫁接苗以及自根苗的抗冷性生理生化指标的变化规律作了研究。结果表明,以棘瓜为砧木的黄瓜嫁接苗叶片的SOD、POD、CAT活性以及Pro、可溶性糖含量均高于后两者,MDA含量低于后两者。说明利用棘瓜嫁接的黄瓜苗可以忍受更长时间或更大强度的低温。