保水剂对水分胁迫下辣椒生长及光合作用的影响

以‘超越5号’辣椒品种为试材,研究了中度水分胁迫10d和20d及随后分别复水的过程中保水剂对始花期辣椒植株的生长量、叶绿素以及光合特性的影响。结果显示:(1)水分胁迫显著抑制了辣椒植株的生长、叶绿素含量以及光合作用。(2)添加2‰的保水剂显著提高了水分胁迫下植株的株高、茎粗、干鲜重、叶绿素含量以及净光合速率、胞间CO2浓度、气孔导度和蒸腾速率等光合参数,降低了气孔限制值和水分利用率。(3)保水剂能够显著缓解短时间内中度水分胁迫造成的伤害,但随着胁迫时间的延长保水剂缓解的效果有限。(4)复水后,添加保水剂的植株生长量、叶绿素含量以及光合参数都显著高于未添加保水剂的植株,同时短时间胁迫下恢复的效果明显好于长时间胁迫的效果。研究表明,施加保水剂能通过提高叶绿素含量,缓解水分胁迫对辣椒叶片气孔的限制,从而提高叶片的光合作用效率,促进辣椒植株的生长,且胁迫时间越短,保水剂缓解的效果越好。     

Application of Vascular Puncture for Evaluation of Curtovirus Resistance in Chile Pepper and Tomato

Curtoviruses cause severe damage to tomatoes and peppers. Functional field resistance to curtoviruses in these plants is desirable but difficult to produce and difficult to screen for because it is time‐consuming and resistance could be achieved by developing resistance either to the virus or to insect feeding. To improve and speed curtovirus resistance testing in tomato (Solanum lycopersicum) and pepper (Capsicum annuum) plants, two puncture methods were developed and compared to leafhopper inoculation and feeding preference assays. The two puncture methods were adapted to introduce a modified Agrobacterium tumefaciens plasmid carrying a recombinant curtovirus into the meristem tissue of tomato plants and into newly germinated chile pepper seedlings. The puncture techniques were used to screen for resistance to curtoviruses in chile pepper and tomato breeding lines and varieties. Similarly, the peppers and tomatoes were assayed for curtovirus resistance using leafhopper inoculation and feeding preference, which was assessed by stylet sheath staining. Virus infection by puncture and leafhopper feeding was monitored using PCR and ELISA. ELISA was performed using an antibody to bacterially expressed coat protein. While pepper cvs Tabasco, NuMex Las Cruces cayenne and New Mexico 6‐4 were infected using both puncture and leafhopper inoculation methods, New Mexico 6‐4 had higher infection rates than the other two cultivars. Stylet sheath staining results suggest that leafhoppers prefer to feed on New Mexico 6‐4 rather than Tabasco and NuMex Las Cruces cayenne. Eight tomato cultivars were infected using meristem removal injection inoculation. Three tomatoes cultivars (CVF‐11, Saladmaster and Supersteak) were infected using leafhopper inoculation, although stylet sheath staining results suggested that the first two cultivars were not preferred by the insect vector. Our results suggest that puncture methods and leafhopper inoculation are successful in resistance screening, and both methods should be used as part of screening, because they assess different types of resistance.     

海南黄灯笼椒与不同品种辣椒的辣椒素含量测定

目的:建立HPLC法测定海南黄灯笼椒等辣椒的辣椒素含量。方法:采用Diamonsil C18柱(4.6 mm×250mm,5μm),流动相为甲醇-水(体积比70∶30),流速为1.0 mL/min,检测波长为230 nm,柱温:30℃,外标法定量。结果:辣椒素的线性关系良好,线性范围2.0μg/mL～20.0μg/mL,r=0.9990;精密度、稳定性实验的RSD均低于2%;平均回收率100.8%(n=9),RSD=1.87%。对海南黄灯笼椒等7个品种辣椒的辣椒素含量进行测定,海南黄灯笼椒的辣椒素含量最高。结论:该方法简便快捷,可用于辣椒中辣椒素含量的测定;海南黄灯笼椒辣椒素含量最高,这些信息可为海南黄灯笼辣椒的研究开发提供参考。     

Thidiazuron induced somatic embryogenesis and plant regeneration in Capsicum annuum

An efficient protocol of direct somatic embryogenesis (without involving intermediate callus) has been developed from stem segments and shoot tips of Capsicum annuum L. Explants were cultured on Murashige and Skoog (MS) medium supplemented with thidiazuron (TDZ). Among the various concentration of TDZ tested, 0.5 μM was proved to be best for induction of somatic embryos. Induction, maturation and germination were achieved on the same medium. The shoots developed from somatic embryos were transferred for rooting to MS medium supplemented with indole-3-butyric acid (IBA). All the regenerated plants with 85 % survival rate were normal with respect to morphology and growth characteristics.     

Induction of plant defence compounds by Pseudomonas chlororaphis PA23 and Bacillus subtilis BSCBE4 in controlling damping-off of hot pepper caused by Pythium aphanidermatum

Bacillus subtilis strain BSCBE4 and Pseudomonas chlororaphis strain PA23 (=P. aureofaciens) were effective biocontrol agents against Pythium aphanidermatum, the causal agent of damping-off of hot pepper (Capsicum annum L.) in greenhouse vegetable production systems. Application of strains BSCBE4 and PA23 at the rate of 20 g kg^-1 of seed significantly increased the growth of hot pepper seedlings. The efficacies of various carriers in sustaining the population of these strains in storage were assessed. Both the antagonists survived up to 180 days of storage in peat and talc-based formulations. The two bacterial strains induced development of plant defence-related enzymes including phenylalanine ammonia lyase, peroxidase, polyphenol oxidase, phenol content, suppressed incidence of damping-off and increased growth of hot pepper seedlings.     

Endophytic bacterial flora in root and stem tissues of black pepper (Piper nigrum L.) genotype: isolation, identification and evaluation against Phytophthora capsici

Aim:  To isolate and identify black pepper ( Piper nigrum L) associated endophytic bacteria antagonistic to Phytophthora capsici causing foot rot disease.
Methods and Results:  Endophytic bacteria (74) were isolated, characterized and evaluated against P. capsici . Six genera belong to Pseudomonas spp (20 strains), Serratia (1 strain), Bacillus spp. (22 strains), Arthrobacter spp. (15 strains), Micrococcus spp. (7 strains), Curtobacterium sp. (1 strain) and eight unidentified strains were isolated from internal tissues of root and stem. Three isolates, IISRBP 35, IISRBP 25 and IISRBP 17 were found effective for Phytophthora suppression in multilevel screening assays which recorded over 70% disease suppression in green house trials. A species closest match (99% similarity) of IISRBP 35 was established as Pseudomonas aeruginosa ( Pseudomonas EF568931), IISRBP 25 as P. putida ( Pseudomonas EF568932), and IISRBP 17 as Bacillus megaterium ( B. megaterium EU071712) based on 16S rDNA sequencing.
Conclusion:  Black pepper associated P. aeruginosa , P. putida and B. megaterium were identified as effective antagonistic endophytes for biological control of Phytophthora foot rot in black pepper.
Significance and Impact of the Study:  This work provides the first evidence for endophytic bacterial diversity in black pepper stem and roots, with biocontrol potential against P. capsici infection.     

辣椒果实中CaRIN转录因子的克隆及其表达分析

根据BLAST获得的表达序列标签设计引物,从辣椒中分离并鉴定了一个新型转录因子,命名为CaRIN。与已知MADS—box蛋白对比表明,CaRIN分别与番茄LeMADS－RIN和矮牵牛FBP4有85％和77％的同源性,并且具有典型的MADS—box类转录因子的特征区域。采用半定量RT－PCR进行特异性表达分析,结果表明CaRIN在破色期和红熟期辣椒果实中大量表达,而在绿熟期辣椒果实和其他辣椒器官中几乎不表达;同时进行了DNA结合位点和系统发生等生物信息学分析,这些结果都表明CaRIN可能参与辣椒果实成熟的调控。     

辣椒雄性不育材料H9A小孢子败育机理

运用光学、电子显微技术及生化指标测定法, 对辣椒（Capsicum annuum）雄性不育材料H9A及其保持系H9B小孢子不同发育时期进行细胞形态学观察及生化特性分析。结果表明, 雄性不育材料H9A的小孢子败育发生在单核前期, 由于绒毡层细胞极度膨胀, 四分体受挤压后破裂并降解, 无法形成正常的单核花粉粒, 属于孢子体败育。不育材料花蕾中的过氧化物酶活性从四分体时期开始明显高于保持系, 保持系游离脯氨酸含量从单核小孢子期开始明显高于不育材料; 不育材料小孢子发育各时期的可溶性蛋白含量都明显低于保持系, 但丙二醛含量均高于保持系。     

辣椒ML基因植物表达载体的构建及其转化

为分析辣椒ML基因在抗辣椒疫病方面的作用,构建了ML基因的植物表达载体pBI121-ML,采用快速冻融法将表达载体导入农杆菌EHA105,并用农杆菌介导法转化辣椒感病品种B12,对得到的转化植株经PCR和RT-PCR分子检测,结果显示,获得了4个辣椒转基因株系.     

Coexpression of a defensin gene and a thionin-like gene via different signal transduction pathways in pepper and Colletotrichum gloeosporioides interactions

The anthracnose fungus, Colletotrichum gloeosporioides, interacts incompatibly with the ripe fruit of pepper (Capsicum annuum). It interacts compatibly with the unripe-mature fruit. We isolated a defensin gene, j1-1, and a thionin-like gene, PepThi, expressed in the incompatible interaction by using an mRNA differential display method. Both genes were developmentally regulated during fruit ripening, organ-specifically regulated, and differentially induced during the compatible and incompatible interactions. Expression of the PepThi gene was rapidly induced in the incompatible-ripe fruit upon fungal infection. The fungus-inducible PepThi gene is highly inducible only in the unripe fruit by salicylic acid. In both ripe and unripe fruit, it was induced by wounding, but not by jasmonic acid. Expression of the j1-1 gene is enhanced by jasmonic acid in the unripe fruit but suppressed in the ripe fruit. These results suggest that both small and cysteine-rich protein genes are induced via different signal transduction pathways during fruit ripening to protect the reproductive organs against biotic and abiotic stresses.