五彩椒(Capsicum annuum L.)的组织培养研究

以抗寒性较强的五彩樱桃椒为试验材料,对观赏椒的组织培养进行了研究。试验表明,最佳外植体为茎尖,茎尖的最佳消毒时间为3min;最佳诱导培养基为MS+6-BA5mg·L-1+NAA1mg·L-1;最佳分化培养基为MS+6-BA5mg·L-1+NAA0.1mg·L-1,最佳生根培养基为1/2MS+IBA0.25mg·L-1。

Effect of Different Culture Conditions on Sporulation Quantity of Phytophthora capsici in Capsicum annuum L. var. dactylus M. in Xunhua

[Objective] The paper was to explore the effect of different culture conditions on sporulation quantity of Phytophthora capsici in Capsicum annuum L.var.dactylus M.in Xunhua.[Method] The effects of light hours,pH,medium and temperature on sporulation quantity during isolation and culture process of P.capsici were studied.[Result] The sporulation quantity of P.capsici under the conditions of 24 h/day light,pH 7.0,potato medium（PSA） and 30 ℃ was the largest,and pH,basal medium and temperature had greater impact on sporulation quantity.[Conclusion]The study laid foundation for the study on natural incidence condition of C.annuum in Xunhua.

气孔在辣椒(Capsicum annuum L.)子叶和下胚轴上的动态变化

以辣椒幼苗(Capsicum annuum L.)为材料,研究了辣椒生长发育过程中子叶的上、下表皮和下胚轴上、下端1 cm处的气孔密度及表皮细胞密度的动态变化。结果表明:(1)辣椒的气孔由肾形保卫细胞组成,无副卫细胞;子叶的表皮细胞呈无规则形,下胚轴的表皮细胞呈长方形或椭圆形;(2)辣椒子叶上、下表皮的气孔密度和表皮细胞密度分别在辣椒种子出芽后第6天和第3天达到最大,之后随子叶的生长逐渐减小,下表皮的气孔密度和表皮细胞密度明显大于上表皮的;(3)辣椒下胚轴上、下端1 cm处的气孔密度随下胚轴的生长变化不明显,但是表皮细胞密度随下胚轴的生长逐渐减小,下胚轴上端的气孔密度和表皮细胞密度明显的大于下胚轴下端的。

Cloning and characterization of CaGID1s and CaGAI in Capsicum annuum L.

Fruit set and development are affected by many phytohormones, including gibberellin. Little is known regarding molecular mechanism underlying gibberellin mediated fruit set and development especially in Capsicum. Three gibberellin recep- tors, CaGIDlb.1, CaGIDlb.2 and CaGIDlc, and a DELLA protein, CaGAI, have been identified in Capsicum annuum L. During the fruit development, the expression level of CaGIDlc was low, and the expression fold change is mild. However, CaGIDlb. 1 and CaGIDlb.2 were relatively higher and more acute, which indicates that CaGIDlb. 1 and CaGID1b.2 may play an important role in fruit pericarp, placenta and seed. Ectopic expressions of CaGIDlb. 1, CaGIDlb.2 and CaGIDlc in Arabidopsis double mutant gidla gidlc increased plant height, among which CaGIDlb.2 had the most significant effect; CaGAI reduced plant height in double mutant rga-24/gai-t6, having a similar function to AtGID1 and AtGAI in stem elon- gation. Yeast two-hybrid （Y2H） and bimolecular fluorescence complementation （BiFC） assays indicated that CaGIDlb.1 and CaGID1 b.2 interact with CaGAI in a GA-dependent manner, while CaGIDlc interacts with CaGAI in a GA-independent manner. Our study reveals the key elements during gibberellin signaling in Capsicum and supports the critical importance of gibberellin for Capsicum fruit set and development.

Combining Ability Analyses of Net Photosynthesis Rate in Pepper (Capsicum annuum L.)

In perspective of breeding high-yield hybrid pepper varieties, combining ability analysis of net photosynthesis rate at different phases of flowering and fruit setting in pepper was made with 15 cross combinations from 6 parents by （1/2） n （n- 1） diallel crosses. There are relatively large differences not only in general combining ability （GCA） effect among different parents and at different phases of flowering and fruit setting, but also in specific combining ability （SCA） effect among different hybrids. There are relatively large GCA effects in late parents but relatively less GCA effects in early parents. No obvious laws have been found in the relationship between SCA effects and maturity of hybrids. Variances of SCA are larger than those of GCA. Heritability is less but influence of environment is larger. Correlation analysis of combining ability between net photosynthesis rate and agronomic character or resistances to main diseases has showed that correlation coefficients of GCA are relatively large at the medium phase and the late phase of flowering and fruit setting. Net photosynthesis rate is more relative to leaf characters and fruit characters. Correlation coefficients of SCA are relatively large at the early phase and the late phase of flowering and fruit setting. Net photosynthesis rate is more relative to leaf characters and plant characters at the early phase but to plant characters and fruit characters at the late phase. Correlation coefficients of SCA between net photosynthesis rate and resistances to main diseases are larger than those of GCA. The combining abilities of net photosynthesis rate at different phases of flowering and fruit setting are positively correlated with those of yield per plant. The combining ability is an important parameter of breeding of high photosynthesis hybrid pepper varieties.

Effect of continuous negative pressure water supply on the growth, development and physiological mechanism of Capsicum annuum L.

Effects of continuous negative pressure water supply on water consumption, growth and development, as well as physiological mechanism and quality of Capsicum annuum L. were investigated in this paper. Meanwhile, the optimal negative pressure water supply conditions for growth of C. annuum L. were screened out to achieve the goals of water conservation, high yield and high quality, thus providing theoretical foundation for its field production. The pot experiment within the greenhouse was utilized; the continuous negative pressure water supply was adopted; the four treatments, artificial watering（CK）, –5 k Pa（T1）, –10 k Pa（T2）, and –15 k Pa（T3） were set; and the daily water consumption, yield, as well as the biomass, nitrate reductase, root activity, vitamin C, capsaicin, and nutrient uptakes of nitrogen（N）, phosphorus（P） and potassium（K） during various stages of its growth were determined. Compared with CK, when the water supply pressure was controlled at –5 to –15 k Pa in the experiment, the total water consumption of C. annuum L. reduced by 53.42 to 67.75%, the total water consumption intensity reduced by 54.29 to 67.14%, and the water use efficiency increased by 12.66 to 124.67%. The N accumulation in a single strain of C. annuum L. from the color turning stage to the red ripe stage increased by 15.99 to 100.55%, respectively, compared with that of CK; the P accumulation increased by 20.47 to 154.00% relative to that of CK, and the K accumulation increased by 64.92 to 144.9% compared with that of CK. Compared with CK, C. annuum L. yield was remarkably improved by 13.79% at T1, and contents of vitamin C, capsaicin as well as carotenoids at all growth stages were enhanced by 13.42–147.01%, 11.54–71.01%, and 41.1–568.06%, respectively. Nitrate reductase activity, root activity and chlorophyll（a＋b） were markedly increased by 335.78–500%, 79.6–140.68% and 114.95–676.19%, respectively, from immature stage to full ripe stage. Adopting the continuous negative pressure water supply for C. annuum L. has a significant water-saving effect, and the water supply pressure being stable at –5 k Pa contributes to its growth and development, improves yield, enhances root activity, promotes nutrient uptake, and improves its quality, thus achieving the effects of water conservation, high yield, high quality and high efficiency.

Combining Transcriptomics and Metabolomics to Uncover the Effects of High-Energy Lithium-Ion Beam Irradiation on Capsicum annuum L.

Hot pepper(Capsicum annuum L.)is consumed as one of the oldest domesticated crops all over the world.Although mutation breeding using radiation has been performed in hot peppers,little is known about the comparative analysis of mutagenic effects at the molecular level by ion beam irradiation.To comprehend the response mechanism of hot pepper to the ion beam,we used a mutant with favorable economic characteristics induced by lithium-ion beam irradiation to investigate the biological effects.The results indicated that the lithium-ion beam had a positive effect on important agronomic traits,particularly yield unit,but had a negligible effect on the photosynthetic rate of hot pepper,with a specific influence on chlorophyll b rather than chlorophyll a.By RNA-Seq analysis,671 up-regulated and 376 down-regulated genes were identified as differentially expressed genes(DEGs)between irradiated and unirradiated hot pepper.Based on GO and KEGG network analysis,the auxin metabolic process was the common pathway in these two networks.A total of 118 potential reactive oxygen species(ROS)scavenging genes and 262 signal transduction genes were identified,suggesting a balance between antioxidant enzymes and enhanced ROS transduction.The amounts of 15 metabolite,involved in GABA pathways,secondary metabolism,carbohydrate metabolism,shikimate pathways,TCA cycles,nitrogen metabolism,glycerol metabolism and acetate pathways,were significantly changed in the ion beam irradiated sample.These results highlighted that the enriched pathways could play important roles in response to ion beam irradiation in hot pepper plants.In summary,these data provide valuable information for future research on ion beam irradiation and genomic studies in hot pepper.

AFLP Analysis of Mitochondrial DNA from a Cytoplasmic Male Sterility Line and its Maintainer in Pepper (Capsicum annuum L.)

Cytoplasmic male sterility (CMS) is a maternally inherited trait characterized by the inability of a plant to produce functional pollen, which is widespread among higher plants. CMS system is a valuable tool for plant

辣椒(Capsicum annuum L.)果实发育及品质相关新microRNA的鉴定与分析

MicroRNA(miRNA)是一种非编码的小RNA,在各种生物过程中起着重要的调节作用。前人研究表明,模式植物中miRNA与果实发育相关。然而,人们对辣椒(Capsicum annuum L.)果实品质及发育相关的miRNA了解尚不清楚。本研究利用二代测序技术(Next-generation sequencing technology)比较了不同品种辣椒果实在不同发育时期的小RNA。利用miRDeep2软件从4个样本中共鉴定出了59个已知miRNA和310个新miRNA。预测了新miRNA的靶基因,共656个,并对其中402个靶基因进行了注释。GO分析和KEGG途径表明,某些靶基因与淀粉糖、氨基糖以及核糖代谢相关。qPCR表明一些miRNA的表达模式与其靶向基因相反,该结果为今后研究miRNA调控果实发育和品质形成提供了重要基础。

Processing of Snack of Goat Chub Chili(Capsicum annuum L.)with Formulations of Tomato and Tomato-Marjoram

The cultivation of goat chili has potentially increased in Chile, due to different varieties of gourmet products made with it;for this reason, new industrial projects are sought to the growing volume to adapt them to the new trends in consumption habits, mainly focused on achieving an attractive meal, a fast preparation and consumption, while nutrients are provided to offer some benefits for human health. The “snacks” products that include the use of fruit satisfactorily respond to these requirements, in addition to providing marketing alternatives to the surplus of fruit exports. For this reason, the present test, consisting of the processing of snacks based on goat chub chili with the addition of tomato and marjoram, was carried out to increase the palatability and the sensorial quality of the final product. The following formulations were used: only chili (T1);chili and tomato (T2);chili, tomato and marjoram (T3). The latter one was characterized by a good flavor and aroma. A large amount of the fruit water content had been removed by drying it with forced hot air. A product without chemical preserves was obtained, with physical and chemical parameters similar to those required by similar foods existing on the market today. In sensorial terms, the formulation to which marjoram was added showed good sensorial and acceptability attributes, highlighting them from the rest of the treatments. The snack color was maintained after the dehydration process, according to panelists.

Purification and Biochemical Characterization of a Protease Inhibitor Ⅱ Family from Jalapeno Pepper(Capsicum annuum L.)

Capsicum annuum L. was initially domesticated in Mexico and northern Central America, and represented an ancient Neotropical plant food complex. The purpose of this paper is to report the isolation and purification of a novo-member of a protease inhibitor from jalape&ntildeo pepper (Capsicum annuum L.) (PIJP). The molecular weight of PIJP inhibitor is 5.95 kDa with 56 amino acids and 6 Cys residues with high inhibitory activity to trypsin with a Ki value of 95 nM. This inhibitor according to the alignment with homologous from NCBI and Pfam databases is a member of proteinase inhibitors II. It is worthwhile to mention a major compositional difference between the proteinase inhibitor II families which have 8 Cys residues. PIJP is the first purified proteinase inhibitor, member of this family with only 6 Cys residues.

几丁质和flg22诱导的辣椒幼苗先天免疫生理响应特征

【目的】为明确几丁质和鞭毛蛋白衍生肽flg22诱导的辣椒幼苗先天免疫生理响应特征,探讨辣椒先天免疫生理响应与辣椒抗多种病害的关系。【方法】以5个四川本地辣椒品种幼苗为试材,鉴定它们对青枯病和疫病病情指数和抗性水平,采用水培法培育幼苗并进行外源几丁质和flg22处理,检测各品种不同诱导时间下幼苗根系生长、气孔孔径、胼胝质沉积、活性氧(ROS)积累和SOD、CAT活性,以及先天免疫相关基因表达量的变化,综合评价其生理响应及其与抗病性关系。【结果】(1)各品种幼苗青枯病和疫病病情指数以‘川腾10号’最低,抗病性最强,以‘本地条椒’最高,抗病性最弱。(2)外源几丁质和flg22抑制了各品种幼苗根系生长速率,诱导离体叶片气孔闭合,促进叶片细胞壁胼胝质沉积加厚,ROS含量持续增加,SOD和CAT活性不断提高。各品种幼苗先天免疫生理响应指标的平均隶属函数值以‘川腾10号’最高,‘本地条椒’最低,并且平均隶属函数值与疫病、青枯病病情指数均具有显著负相关性。(3)外源flg22和几丁质诱导‘川腾10号’幼苗先天免疫相关基因CaWRKY22、CaMAPK7和ChiIV3显著上调表达。【结论】外源flg22和几丁质可诱导辣椒幼苗先天免疫生理响应,且响应程度强弱在品种间存在差异,依据生理响应指标通过隶属函数可综合评价辣椒品种的抗病水平;‘川腾10号’的平均隶属函数值最高,多抗性水平最好,这与其先天免疫相关基因CaWRKY22、CaMAPK7和ChiIV3的显著上调表达有关。

辣椒EIL基因家族的鉴定及其在盐碱胁迫下的表达分析

EIL(Ethylene-insensitive 3-like)基因在参与植物乙烯信号通路的转导和生长发育过程中发挥着重要作用。为了解辣椒EIL基因家族成员信息,利用生物信息学手段对辣椒EIL基因家族成员的理化性质、蛋白质结构、系统进化、基因结构、保守域、启动子顺式作用元件及CaEILs基因在不同组织部位和不同非生物胁迫条件下的表达模式进行分析。同时使用实时荧光定量PCR(qRT-PCR)对辣椒叶片中CaEILs在盐碱胁迫下的表达模式进行研究。结果表明,辣椒9个CaEILs分布在6条染色体上,氨基酸数目、蛋白质理论分子质量和脂肪系数分别介于209~677、23.77~76.07 ku和63.10~87.75,且主要为酸性、亲水的不稳定性核蛋白。系统进化分析显示,CaEILs被分成了4个亚家组,9个CaEILs在根、茎、叶、花蕾、花中具有不同程度的表达,而低温、高温、高盐、干旱胁迫下均能诱导CaEILs的表达,其对以上非生物胁迫均具有不同程度的响应。此外,利用qRT-PCR对盐碱胁迫下辣椒叶片中CaEILs进行了检测,结果发现,随着处理时间的延长,CaEIL1、CaEIL2、CaEIL4、CaEIL5、CaEIL8的表达量整体呈上升趋势,而CaEIL3、CaEIL6、CaEIL7、CaEIL9的表达量整体呈下降趋势。

辣椒分子遗传图谱的构建和胞质雄性不育恢复性的QTL分析

以辣椒胞质雄性不育的保持系Yolowonder、恢复系Perennial及其F1构建的DH群体与不育系 770 13A测交的群体为供试材料 ,用AFLP分子标记技术构建了包括 43个标记 8个连锁群的辣椒分子遗传图谱。在对测交群体育性调查的基础上 ,把控制辣椒胞质雄性不育恢复性的QTL初步定位到第 1、3、6、8个连锁群上。

In vitro Plant Regeneration of Pepper Cytoplasmic Male Sterility (CMS) Lines via Cotyledon Culture

An in vitro shoot regeneration procedure was developed in pepper （ Capsicum annuum L. ） cytoplasmic male sterility （CMS） lines 9704A and 8214A using cotyledon as explant. The callus and bud cluster derived from cotyledon tissue explants were proliferated on Murashige and Skoog （MS） medium supplemented with different combinations of 6-benzladenine （6-BA）, indole-3-acetic acid （IAA）, gibberellic acid （GA3） and silver nitrate （AgNO3）. From the formula of MS appended with 5.0 mg/L 6-BA, 1.0 mg/L IAA and 5.0 mg/L AgNO3, for the explants callus and bud cluster, the maximum differentiation rates （ respectively 100.0% and 58.3% ） and average number of adventitious bud from each explant （respectively 18.8 and 13.2） were obtained. The optimum medium combination for the elongation of adventitious bud was determined to be： MS ＋ 3.0 mg/L 6-BA ＋ 1.0 mg/L IAA ＋ 5.0 mg/L AgNO3 ＋ 2.0 mg/L GA3, from which the elongation rates of buds from callus and bud cluster were both 100%, and the average number of per explant adventitious bud number reached 6.3 and 5.8, respectively. And all the elongated shoots were successfully rooted on half-strength MS medium supplemented with 0.3-0.5 mg/L IAA.

利用分子标记对辣椒抗马铃薯Y病毒的3个QTLs进行选择

为获得含有特定基因型的个体,深入研究辣椒抗马铃薯Y病毒(PVY)不同QTLs之间的关系,利用AFLP、SCAR和CAPS标记在辣椒材料‘Perennial’和‘Yolo Wonder’组成的BC_2和BC_1S_1分离世代对抗PVY的3个QTLs位点进行了选择。结果从BC_2群体中选出47个在主效QTL上含有抗性位点的单株,从中选出25个在LG9 PY2染色体上含有抗性位点的单株,最后选出11个在LG11 P10染色体上含有抗性位点的单株;从BC_1S_1群体中选出55个在LG9 PY2染色体上具有纯合抗性位点的单株,从中选出12个在主效QTL上纯合抗性位点的单株。经卡平方检测,实际分离比例同理论分离比例差异不显著。

用AFLP技术分离辣椒mtDNA中与雄性不育相关的基因片段

采用AFLP技术对辣椒胞质雄性不育系‘北A’、相应保持系‘北B’及其杂种F1的m tDNA进行了比较分析,获得了9条差异谱带,对全部差异谱带进行了克隆测序和同源性比对分析。结果表明,部分序列同烟草NADH脱氢酶亚基1和核糖体蛋白L2基因、矮牵牛线粒体嵌合基因nad4L-orf25等核苷酸序列存在90%以上的相似性。推断这些差异片段可能与辣椒细胞质雄性不育相关。

辣椒细胞质雄性不育基因的AFLP分析

本文采用AFLP(amplified fragment length polymorphism)技术对辣椒(Capsicum annuum L.)细胞质雄性不育系21A及其相应的保持系21B基因组DNA进行多态性比较,筛选了64对AFLP选择性引物PstⅠ-NNN+MseⅠ-NNN组合,有60对引物组合均在两系间扩增出5447条带,多态性位点为4个,其中在不育系材料中仅有2个多态性片段AGC/CGC378和ATC/GAG446,对特异片段进行克隆、测序,其序列全长分别为340bp和408bp,它们编码的氨基酸序列与烟草基因组线粒体DNA序列同源性达94%和96%。

HPLC测定辣椒中Nonivamide含量

建立高效液相色谱法测定辣椒样品中Nonivamide(合成辣椒碱)的方法。样品用甲醇-四氢呋喃(1∶1)混合溶液超声提取,以甲醇-水(60∶40,其中每升流动相中含25mmol硝酸银和2.5mL冰醋酸)为流动相,在280nm处测定。线性方程为Y=7180.5C+215.6(r=0.9995),Nonivamide在0.28μg/mL～27.68μg/mL范围内呈良好的线性关系,回收率为96.72%。在天然生长的辣椒中含有Nonivamide,但其含量未超过辣椒素类物质总量的2.0%。

辣椒AFLP反应体系的优化与建立

以5个辣椒(Capsicum annuum L.)材料为研究对象,对AFLP反应体系中的DNA用量、酶切连接时间、预扩增产物的稀释倍数等关键因素进行优化分析,建立了适宜辣椒作物的AFLP反应体系。研究结果:酶切连接反应中,基因组DNA适宜用量为100ng,反应时间是6h最为合适,预扩增产物适宜稀释倍数在30～50倍时较为理想。