Geraniol and linalool synthases from wild species of perilla

Geraniol and linalool synthases were isolated from three pure strains of Perilla hirtella and Perilla setoyensis, which are wild species of perilla. Their amino acid sequences were very similar to those of Perilla citriodora and Perilla frutescens that were reported previously. However, comparison of the sequences of the same functional synthases derived from different species of Perilla demonstrated that the similarities were high among P. citriodora, P. hirtella and P. frutescens, but low between P. setoyensis and any of the others. This result corresponds well with our previous results showing that P. setoyensis is remotely related to the other perilla species. Both geraniol and linalool synthases utilize geranyl diphosphate (GDP) as their catalytic substrate and they were expressed simultaneously in perilla. The linalool synthase is considered to be the enzyme whose metabolite seems not to be oxidized nor reduced in the plant body and the geraniol and limonene synthases are the initial-step-catalyzing enzymes for a variety of oil compounds. The regulation of the substrate flow between them would be interesting for further study.     

模拟氮沉降对紫苏叶挥发油主要成分的影响

我国为世界三大高氮沉降区之一,氮沉降严重影响了植物生长发育。该研究采用喷施硝酸铵(NH4NO3)模拟氮沉降,分析了不同浓度氮沉降作用下紫苏叶中紫苏醛、D-柠檬烯、α-蒎烯等3种挥发油成分的变化规律。结果表明:随喷施氮盐浓度不断提高,紫苏叶挥发油的3种主要成分含量均有显著下降趋势;氮盐浓度升至0.044 mol·L~-1时,紫苏醛、D-柠檬烯、α-蒎烯的含量降至最低,之后趋于稳定;氮盐浓度对3种挥发油成分含量的比例也有影响;不同氮盐浓度处理下,3种挥发油成分的变异系数不同,紫苏醛的变异系数为0.692 9,D-柠檬烯的变异系数为0.460 1,而α-蒎烯的变异系数为0.271 6,即紫苏醛含量变化最大,α-蒎烯含量最为稳定。大气氮沉降浓度对紫苏叶挥发油主要成分含量有显著影响,随氮盐浓度不断提高,紫苏醛、D-柠檬烯、α-蒎烯等3种挥发油成分含量呈降低趋势,尤以紫苏醛含量的降低最为剧烈。氮沉降增加对紫苏叶有效成分含量有降低的作用。     

紫苏属植物染色体数目和核型分析

通过对26份紫苏属材料进行染色体数目的观察,并对其1种(包括紫苏和白苏)及2变种材料各一份进行核型分析.结果显示,紫苏属植物染色体数均为2n=40,其核型公式分别为紫苏[P.frutescens(L.)Britton var.frutescens(zi-su)]2n=40=20m(2SAT) 10sm 10st;野生紫苏[P.frutescens var.purpurascens(Hayata)H.W.Li]2n=40=18m 14sm(2SAT) 8st;回回苏[P.frutescens var.crispa(Benth.)Deane ex Bailey]2n=40=10m(2SAT) 14sm 16st;白苏[P.frutescens(L.)Britton var.frutescens(bai-su)]2n=40-24m 12sm(2SAT) 4st.结果表明,紫苏属不同原(变)种植物的染色体数日相同,但在核型上存在一定的差异,其染色体核型为首次报道.     

Essential oil variation of cultivated and wild Perilla analyzed by GC/MS

Twenty components extracted from the essential oil in the leaves of 172 samples of Perilla frutescens var. crispa (vegetable crop form), P. frutescens var. frutescens (oil crop form), the wild/weedy form of P. frutescens, and three wild Perilla species, Perilla citriodora, Perilla hirtella and Perilla setoyensis were analyzed using GC/MS. A wide range of essential oil components were found among the wild/weedy form of P. frutescens, whereas distinctive components were detected in each wild Perilla species. Egomaketone, asaron, methyleugenol and 4,6-dimethoxy- or 4,7-dimethoxy-5-(2-propenyl)-1,3-dioxaindan were detected from Perilla for the first time. Limonene derivatives, piperitone and piperitenone, were detected from P. citriodora. Discovery of the limonene derivatives in this Perilla species provides evidence of this wild species being a genome donor of P. frutescens, while limonene synthase has been considered to be a specific enzyme in cultivated P. frutescens. These results will be useful for the evaluation and utilization of Perilla genetic resources.     

Lipid biosynthesis in developing perilla seeds

In developing seeds of Perilla frutescens var. crispa, the triacylglycerol fraction was found to accumulate between 15 and 19 days after flowering. Of this, 65% of the total fatty acids was alpha-linolenic acid in the mature seeds, with the latter being esterified in comparable amounts at all positions (sn-1, 2 and 3) of the glycerol residue. It was also demonstrated that, 1-acylglycerol-3-phosphate acyltransferase, which catalyzes esterification at the sn-2 position of the glycerol backbone, showed low activities for alpha-linolenoyl-CoA as substrate. These findings suggest that the diacylglycerol precursor for triacylglycerol synthesis is not directly derived from phosphatidic acid through the glycerol phosphate pathway.     

紫苏硬脂酰-ACP脱氢酶基因家族鉴定及表达分析北大核心CSCD

硬脂酰-ACP脱氢酶(SAD)催化硬脂酸脱氢生成油酸,是形成不饱和脂肪酸的关键酶。该研究从紫苏转录组数据库中筛选鉴定紫苏硬脂酰-ACP脱氢酶(PfSAD)家族基因,并进行生物信息学分析及保守功能域分析,用qRT-PCR技术检测PfSADs各成员在不同组织中的表达特性,以探讨PfSAD家族基因在调控种子脂肪酸组分中的作用,为紫苏脂肪酸组分的遗传改良提供基因元件。结果显示:(1)从该课题组前期自测的紫苏转录组数据库中共检测出6个PfSAD家族基因,其编码蛋白的氨基酸长度介于367~396 aa之间,均具有SAD的保守结构域和二铁中心,预测其基因编码蛋白均定位于叶绿体。(2)多序列比对结果显示,紫苏PfSADs蛋白序列与拟南芥、蓖麻及可可等植物的SAD蛋白序列相似性均在50%以上;系统进化分析显示,6个紫苏SAD蛋白被分为3个亚组,其中第一个亚组包含PfSAD1,第二亚组包含PfSAD2、PfSAD3,第三亚组包含PfSAD4、PfSAD5和PfSAD6。(3)实时荧光定量PCR分析发现,PfSADs各成员在‘晋紫苏1号’不同组织中的表达量差异显著,其中PfSAD1主要在叶中表达,PfSAD2、PfSAD3、PfSAD4和PfSAD5在种子中表达量较高,PfSAD6在花中具有显著表达优势。研究表明,PfSADs具有典型的保守基序及催化SAD的活性中心,其各成员在不同的组织中高表达,推测这6个基因均参与了硬脂酰ACP(C18∶0-ACP)脱氢生成油酰基ACP(Δ9C18∶1-ACP)的过程,在紫苏油脂合成代谢过程中发挥重要作用。     

高温高湿胁迫下茉莉酸甲酯对紫苏种子萌发及生理特性的影响

以紫苏品系GS011种子为材料,在温度为50℃、空气相对湿度为95%、时间为12h的胁迫条件下,研究不同浓度(0.05、0.10、0.15、0.20和0.25mmol.L-1)茉莉酸甲酯(MeJA)对紫苏种子萌发、幼苗生长、叶片氧化损伤和抗氧化酶活性的影响。结果表明:0.15mmol.L-1 MeJA处理能显著缓解高温高湿对紫苏种子生长发育造成的胁迫伤害,使紫苏种子发芽率、发芽势、发芽指数、活力指数均达到最大值,分别为40.84%、31.83%、8.83和1.35;同时使其叶片MDA含量最低(0.15μmol.g-1),SOD、CAT、LOX、POD和APX活性均达到最大值,分别为0.27U.mg-1及4.7、5.7、4.9和8.9U.g-1.min-1。研究发现,MeJA在一定程度上能提高SOD等保护酶的活性,缓解高温高湿造成的氧化损伤,有效促进高温高湿胁迫下紫苏种子的萌发和幼苗生长。     

A constitutively expressed Myc-like gene involved in anthocyanin biosynthesis from Perilla frutescens: molecular characterization,heterologous expression in transgenic plants and transactivation in yeast cells

The coordinate expression of anthocyanin biosynthetic genes in leaves and stems of a red forma of Perilla frutescens is presumably controlled by regulatory gene(s). A Myc-like gene (Myc-rp) was isolated from a cDNA library prepared from the leaves of red P. frutescens, and its deduced amino acid sequence shows 64% identity with that of delila from snapdragon. The Myc-rp gene was expressed in leaves and roots of both red and green P. frutescens equally. Comparison of deduced amino acid sequence of Myc-rp with that of Myc-gp, the second allele isolated from a green forma of P. frutescens, indicates that the 132nd amino acid, alanine, existing in MYC-RP was changed to serine in MYC-GP. The heterologous expression of these two alleles of Myc-like gene in tobacco and tomato resulted in an increase of the anthocyanin contents in flowers of tobacco and vegetative tissues and flowers of tomato. However, the flowers of transgenic tobacco expressing the fragment with a partial deletion (encoding 1–115 amino acids deleted) of Myc-gp gave no change in anthocyanin accumulation, but some morphological changes of the flower were observed. In yeast, the MYC-RP/GP and Delila protein exhibited transactivation activity on the GAL-1 promoter from yeast and the promoter of dihydroflavonol 4-reductase (DFR) gene from P. frutescens. A transactivation domain of MYC-RP/GP and Delila could be located in the region between the 193rd and the 420th amino acid of MYC-RP/GP proteins. Our data indicate that this Myc-like gene presumably functions in the regulation of anthocyanin biosynthesis similarly in different tissues of dicot plants.