Ethylene biosynthesis in Amaranthus caudatus seeds in response to methyl jasmonate

Methyl jasmonate (JA-Me) at 10^–3 M completely inhibited Amaranthus caudatus seed germination. Exogenous ethylene could totally reverse this inhibition. The inhibitor of ethylene action, 2,5-norbornadiene (NBD), increased the sensitivity of seeds to JA-Me. Methyl jasmonate inhibited ethylene production and also decreased both 1-aminocyclopropane-1-carboxylic acid (ACC) and malonyl ACC (MACC) content. Likewise, ACC oxidase activity in vivo was decreased by jasmonate. Similarly ACC oxidase activity in vitro isolated from seeds incubated in the presence of JA-Me was lower than that isolated from untreated seeds.The inhibitory JA-Me action on seed germination seems to be mainly associated with the inhibition of ethylene biosynthesis. Both inhibition of ACC synthase and ACC oxidase activity and/or synthesis can be involved.     

Different fermentation strategies by Schizochytrium mangrovei strain pq6 to produce feedstock for exploitation of squalene and omega‐3 fatty acids

Schizochytrium mangrovei strain PQ 6 was investigated for coproduction of docosahexaenoic acid (C22: 6ω‐3, DHA ) and squalene using a 30‐L bioreactor with a working volume of 15 L under various batch and fed‐batch fermentation process regimes. The fed‐batch process was a more efficient cultivation strategy for achieving higher biomass production rich in DHA and squalene. The final biomass, total lipid, unsaponifiable lipid content, and DHA productivity were 105.25 g · L^?1, 43.40% of dry cell weight, 8.58% total lipid, and 61.66 mg · g^?1 · L^?1, respectively, after a 96 h fed‐batch fermentation. The squalene content was highest at 48 h after feeding glucose (98.07 mg · g^?1 of lipid). Differences in lipid accumulation during fermentation were correlated with changes in ultrastructure using transmission electron microscopy and Nile Red staining of cells. The results may be of relevance to industrial‐scale coproduction of DHA and squalene in heterotrophic marine microalgae such as Schizochytrium .     

季节变化对聚果榕-榕小蜂互利共生系统生长与繁殖的影响

榕属植物及其传粉昆虫榕小蜂是自然界协同进化的经典模型,榕果内雌花资源如何分配一直是备受关注的问题。为验证季节变化对榕树-榕小蜂互利共生系统生长与繁殖的影响,该研究以西双版纳地区的聚果榕(Ficus racemosa)为材料,分析了季节变化对榕果大小、自然进蜂量以及榕树-榕小蜂繁殖的影响,并利用人工控制性放蜂实验和模型拟合,探讨榕果最适进蜂量及不同季节进蜂量对雌花资源分配的影响。结果表明:季节对榕果直径有显著影响,雨季的榕果直径显著小于干热季和雾凉季;不同季节的自然进蜂量也有显著差别,苞片口对调节进蜂数量有重要作用;季节对榕树-榕小蜂繁殖分配也有影响,雾凉季产生的种子数量和榕小蜂数量均最多;同时人工控制实验和二次抛物线模型拟合结果表明,母代雌蜂数量与种子及榕小蜂后代数量均呈抛物线关系,雌蜂数量过多或过少都对榕树-榕小蜂的繁殖不利,自然进蜂量与拟合的最优进蜂量基本一致。研究结果说明榕果进化出了适应西双版纳地区季节变化的繁殖策略。     

Up-regulation of soyasaponin biosynthesis by methyl jasmonate in cultured cells of Glycyrrhiza glabra

Exogenously applied methyl jasmonate (MeJA) stimulated soyasaponin biosynthesis in cultured cells of Glycyrrhiza glabra (common licorice). mRNA level and enzyme activity of beta-amyrin synthase (bAS), an oxidosqualene cyclase (OSC) situated at the branching point for oleanane-type triterpene saponin biosynthesis, were up-regulated by MeJA, whereas those of cycloartenol synthase, an OSC involved in sterol biosynthesis, were relatively constant. Two mRNAs of squalene synthase (SQS), an enzyme common to both triterpene and sterol biosyntheses, were also up-regulated by MeJA. In addition, enzyme activity of UDP-glucuronic acid: soyasapogenol B glucuronosyltransferase, an enzyme situated at a later step of soyasaponin biosynthesis, was also up-regulated by MeJA. Accumulations of bAS and two SQS mRNAs were not transient but lasted for 7 d after exposure to MeJA, resulting in the high-level accumulation (more than 2% of dry weight cells) of soyasaponins in cultured licorice cells. In contrast, bAS and SQS mRNAs were coordinately down-regulated by yeast extract, and mRNA accumulation of polyketide reductase, an enzyme involved in 5-deoxyflavonoid biosynthesis in cultured licorice cells, was induced transiently by yeast extract and MeJA, respectively.     

Stimulation of Rg3 ginsenoside biosynthesis in ginseng hairy roots elicited by methyl jasmonate

It has been recognized that ginsenoside Rg3 is not naturally produced in ginseng although this ginsenoside can accumulate in red ginseng as the result of a thermal process. In order to determine whether or not Rg3 is synthesized in ginseng, hairy roots were treated with methyl jasmonate (MJ). From HPLC analysis, no peak for Rg3 was observed in the controls. However, Rg3 did accumulate in hairy roots that were MJ-treated for 7?days. Rg3 content was 0.42?mg/g (dry weight). To gain more insight into the effects of MJ on UDP-glucosyltransferase (UGT) activity, we attempted to evaluate ginsenoside Rg3 biosynthesis by UGT. A new peak for putative Rg3 was observed, which was confirmed by LC-MS/MS analysis. Our findings indicate that the proteins extracted from our hairy root lines can catalyze Rg3 from Rh2. This suggests that our ginseng hairy root lines possess Rg3 biosynthesis capacity.     

Induction of volatile terpene biosynthesis and diurnal emission by methyl jasmonate in foliage of Norway spruce

Terpenoids are characteristic constitutive and inducible defense chemicals of conifers. The biochemical regulation of terpene formation, accumulation, and release from conifer needles was studied in Norway spruce [Picea abies L. (Karst)] saplings using methyl jasmonate (MeJA) to induce defensive responses without inflicting physical damage to terpene storage structures. MeJA treatment caused a 2-fold increase in monoterpene and sesquiterpene accumulation in needles without changes in terpene composition, much less than the 10- and 40-fold increases in monoterpenes and diterpenes, respectively, observed in wood tissue after MeJA treatment (D. Martin, D. Tholl, J. Gershenzon, J. Bohlmann [2002] Plant Physiol 129: 1003-1018). At the same time, MeJA triggered a 5-fold increase in total terpene emission from foliage, with a shift in composition to a blend dominated by oxygenated monoterpenes (e.g. linalool) and sesquiterpenes [e.g. (E)-beta-farnesene] that also included methyl salicylate. The rate of linalool emission increased more than 100-fold and that of sesquiterpenes increased more than 30-fold. Emission of these compounds followed a pronounced diurnal rhythm with the maximum amount released during the light period. The major MeJA-induced volatile terpenes appear to be synthesized de novo after treatment, rather than being released from stored terpene pools, because they are almost completely absent from needle oleoresin and are the major products of terpene synthase activity measured after MeJA treatment. Based on precedents in other species, the induced emission of terpenes from Norway spruce foliage may have ecological and physiological significance.     

Overexpression of acetyl-CoA synthetase increased the biomass and fatty acid proportion in microalga Schizochytrium

High acetate accumulation was produced during glucose fermentation in high cell density cultures, which is harmful to cell growth. In order to reduce the negative impact of acetate accumulation on the fermentation products, we introduced the Escherichia coli acetyl-CoA synthetase (ACS) gene into the marine microalga Schizochytrium sp. TIO1101, generating genetically modified ACS transformants. The results of PCR and blotting analyses showed that the exogenous ACS gene was incorporated into the genome and successfully expressed. The engineered Schizochytrium increased the pH value and reduced the acetate concentration in the final fermentation medium significantly. Furthermore, the ACS transformants exhibited faster growth and glucose consumption rates than the wild-type strain. The biomass and fatty acid proportion of ACS transformants increased by 29.9 and 11.3 %, respectively. Taken together, the data suggest that ACS overexpression in Schizochytrium might improve the utilization of carbon resource and decrease the production of acetate byproduct. These results demonstrate that application of ACS in metabolic genetic engineering could improve the properties of Schizochytrium significantly.     

Enhancement of ginsenoside biosynthesis in high-density cultivation of Panax notoginseng cells by various strategies of methyl jasmonate elicitation

A single addition of 200 M methyl jasmonate (MJA) to high-density cell cultures of Panax notoginseng enhanced ginsenoside production in both shake-flask (250 ml) and airlift bioreactor (ALR; 1 l working volume). Repeated elicitation with two additions of 200 M MJA during cultivation further induced the ginsenoside biosynthesis in both cultivation vessels. The content of ginsenosides Rg₁, Re, Rb₁ and Rd in the ALR was increased from, respectively, 0.18±0.01, 0.21±0.01, 0.21±0.02 and 0 mg per100 mg dry cell weight (DW) in untreated cell cultures (control) to 0.32±0.02, 0.36±0.02, 0.72±0.06 and 0.08±0.01 mg per100 mg DW with a single addition of MJA and further increased to 0.43±0.02, 0.46±0.03, 1.09±0.07 and 0.14±0.02 mg per100 mg DW with two additions of MJA. Interestingly, the activity of the Rb₁ biosynthetic enzyme (UDPG-ginsenoside Rd glucosyltransferase), was also increased with a single elicitation by MJA and increased again by a repeated elicitation, which coincided well with the trend in the increase in Rb₁ content. In order to further improve the cell density and ginsenoside production, a strategy of MJA repeated elicitation combined with sucrose feeding was adopted. The final cell density and total ginsenoside content in the ALR reached 27.3±1.5 g/l and 2.02±0.06 mg per100 mg DW; and the maximum production of ginsenoside Rg₁, Re, Rb₁ and Rd was 111.8±4.7, 117.2±4.6, 290.2±5.1 and 32.7±8.1 mg/l, respectively. The strategies demonstrated and the information obtained in this work are useful for the efficient large-scale production of bioactive ginsenosides by plant cell cultures.     

茉莉酮酸甲酯对水稻化感物质的诱导效应

在室内和田间条件下 ,外源茉莉酮酸甲酯均能显著地诱导水稻化感物质的合成 ,而且这种诱导效应与施用茉莉酮酸甲酯的浓度和诱导时间显著相关。 0 .4 m mol/L浓度和处理后 4 8h,茉莉酮酸甲酯对水稻化感物质的诱导效应最强。同样 ,不同的水稻品种对茉莉酮酸甲酯的诱导响应也有显著差异。水稻化感品种 PI312 777和丰华占在茉莉酮酸甲酯的诱导下能很快合成大量的化感物质 ,而水稻非化感品种华粳籼的化感物质的含量虽也有所增加 ,但达不到能显示化感作用的浓度。进一步实验证明 :茉莉酮酸甲酯在处理 4 8h后虽能诱导水稻品种合成大量的化感物质 ,但这一诱导效应并不能长期维持。研究揭示 :水稻化感物质的合成可在外部因子的作用下动态变化 ,这对揭示和充分利用水稻的化感作用机制有重要意义。     

外源茉莉酸甲酯对辣椒烟粉虱的影响

化学农药仍是目前蔬菜烟粉虱Bemisia tabaci防控的重要措施,研发有效的非化学措施防控烟粉虱是蔬菜绿色生产亟待解决的问题。本文以苏椒15号（感性品种）和新苏椒五号（抗性品种）辣椒Capsicum annuum L.为对象,通过在辣椒上喷施茉莉酸甲酯,研究外源茉莉酸甲酯对烟粉虱发育历期、存活率、消化酶活性和田间烟粉虱种群动态的影响。研究发现,取食茉莉酸甲酯处理的辣椒后烟粉虱的发育历期显著延长、存活率明显下降。其中取食感性品种辣椒和抗性品种辣椒的烟粉虱发育历期分别延长7.66 d和9.66 d,存活率分别下降10.5%和9.89%;取食感性品种辣椒的烟粉虱主要是延长卵期、4龄若虫期和伪蛹期,而取食抗性品种辣椒的烟粉虱各历期均延长。外源茉莉酸甲酯对烟粉虱体内消化酶的活性也有明显的影响,取食茉莉酸甲酯处理的辣椒后烟粉虱的消化酶活性显著下降,取食感性品种辣椒和抗性品种辣椒24 h后,胰蛋白酶活力分别较对照下降了61.86%和45.0%,糜蛋白酶活性分别较对照下降了47.29%和40.11%。大田辣椒上喷施茉莉酸甲酯后,烟粉虱的种群数量迅速下降,但在抗性品种和感性品种上烟粉虱的校正虫口减退率没有明显的差异。研究发现,外源茉莉酸甲酯处理,可以延缓烟粉虱的发育速率,降低胰蛋白酶和糜蛋白酶的活性,对田间烟粉虱种群也有较好的控制作用。