Inhibition of polygalacturonase and polygalacturonic acid lyase fromErwinia carotovora subsp.carotovora by phenolics in vitro

Summary The phenolic acids benzoic, caffeic, chlorogenic, ferulic, p-coumaric, protocatechuic, salicylic, sinapic, syringic and vanillic together with vanillin, were tested for their ability to inhibit polygalacturonic acid lyase (PL) and polygalacturonase (PG) in culture filtrates ofErwinia carotovora subsp.carotovora. None of the compounds inhibited PL at 200 μg/ml, although syringic and sinapic acids caused a 54% and 43% reduction respectively at 400 μg/ml. PG was strongly inhibited by caffeic acid (57% inhibition at 100 μg/ml) and chlorogenic acid caused slight inhibition. Little if any inhibition of PG was caused by benzoic, ferulic, p-coumaric, protocatechuic, salicylic, sinapic, syringic, and vanillic acids or vanillin. The percentage inhibition of PG by chlorogenic acid was not affected by pH values over the range 4–7.     

The involvement of phenolics and phytoalexins in resistance of potato to soft rot

Summary Phenolics, for example chlorogenic, caffeic and ferulic acid, and phytoalexins, such as rishitin and phytuberin, were identified in potato tubers cv. Kufri Chandramukhi. The tissue of healthy tubers contained no detectable phytoalexins but did contain phenolics. The levels of these compounds were correlated with soft rot development. The rotting tissue either was free of these groups of compounds or had low concentrations. The wound periderm formed as a result of recovery from injury and infection contained high levels of the compounds. Much higher concentrations were detected at lower storage temperatures when oxygen supply was adequate. Antibacterial properties of the phenolics identified were tested againstErwinia carotovora which was inhibited by chlorogenic, caffeic and ferulic acids. The three phenolics were more effective together, in proportions in which they occurred in wound periderm, than individually. It was observed that none of these phenolics could inhibit pectolytic enzymes ofE. carotovora.     

4CL基因在海巴戟叶片莨菪亭累积过程中的功能研究

为明确海巴戟叶片中莨菪亭累积规律,探究4CL基因在其累积过程中起到的作用。利用高效液相色谱仪对离体的海巴戟成熟叶0～48 h内莨菪亭含量进行测定,并测定经不同浓度的阿魏酸处理后莨菪亭含量的变化。以阿魏酸为底物,利用紫外分光光度计进行酶活反应的4CL总酶活性测定。对RNA-seq结果中7个4CL基因进行qPCR,筛选出一个表达趋势与莨菪亭含量及酶活变化趋势一致的4CL基因作为关键候选基因,进行全长克隆以及生物信息学分析,最后用qRT-PCR研究其表达特性。海巴戟叶片在采摘后48 h内,莨菪亭含量在12 h时最高,达0.35 mg/g,48 h最低,为0.18mg/g。4CL总酶活性变化与莨菪亭含量趋势相近。阿魏酸处理后的叶片中4CL总酶活增加。通过qRT-PCR筛选到4CL(DN15707)基因,克隆、测序后发现其长度为1632 bp,具备完整开放阅读框,在NCBI进行序列比对及系统进化树分析,确定该基因为4CL基因家族中的4CL2基因,命名为Mc4CL2。Mc4CL2基因可能是莨菪亭累积过程中的关键基因,它能启动4CL酶活,充分利用阿魏酸底物进而导致莨菪亭累积。     

In vitro induction of potato tuberization by organic acids

Summary Potato microtubers are usually induced in media containing growth regulatory, typically cytokinins and growth retardants. However, since these substances may imbalance the physiology of the cultured explants and cause adverse carry-over effects on the subsequent performance of the microtubers, the potential acid-induction of in vitro-mass tuberization was investigated as an alternative. For this purpose, a range of explant types (stolons, single-node or apex-containing multinodal sections), organic acids (acetic, propionic, ascorbic, acetylsalicylic or salicylic acid), and photoperiods (continuous dark, 8 or 16 h photoperiod) were established. Gellified medium or raft membranes on liquid medium were also tested. Although variations due to explant type, photoperiod, organic acid supplemented, and medium type were found, all organic acids tested under the different experiments caused tuberization. Multiple tuber formation or bigger tubers did not develop on multinodal explants. In terms of tuberization rate and mean tuber weight, gellified medium performed better than liquid medium with rafts. The recently established role of the salicylic and acetylsalicylic acids on tuberization is corroborated by our system. The results with the acetic, propionic and ascorbic acids show that in vitro hormone-free tuberization can be easily and rapidly achieved.     

外源水杨酸诱导的部分甜瓜自毒作用相关基因的表达分析

以甜瓜(Cucumis melo)品种‘新银辉’为材料,采取根灌法,以水杨酸(10μmol/L)处理甜瓜幼苗,通过Real-Time PCR对叶片中部分自毒作用相关基因的表达量进行定量分析,以探究外源水杨酸诱导对甜瓜自毒作用的影响。结果显示,经外源水杨酸诱导后:(1)在催化苯丙烷类代谢途径的3个初始反应的蛋白酶编码基因中,PAL和C4H随处理后时间增加,呈逐渐上调趋势,但PAL比C4H更快响应胁迫,而4CL转录水平没有显著变化;(2)在类黄酮生物合成的3个关键酶的基因中,CHI、F3H均显示出不同程度的下调,CHS于处理后1 d略微上调,然后恢复至处理前水平;(3)两个转录因子基因WD40、R2R3-MYB分别在不同处理时间后显著下调;(4)WD40与CHI的表达相关性较高。据上述结果 ,水杨酸诱导后:(Ⅰ)CHI表达可能与转录因子WD40蛋白相关;(Ⅱ)WD40、R2R3-MYB与PAL、C4H、4CL、CHS间不存在简单、明确的调控关系;(Ⅲ)柚皮苷、表儿茶素、芦丁等甜瓜主要化感物质合成减少。     

Alpha-amylase treatment increases extractable phenolics and antioxidant capacity of oat (Avena nuda L.) flour

In this work, α-amylase was used to treat oat flour with the intent to release phenolic compounds with known antioxidant properties. After methanol extraction, the amounts of nine beneficial phenolic compounds were measured using HPLC. The antioxidant activities of the extracts were assessed using 2,2′-azinobis (3- ethylbenzothiazoline-6-sulfonic acid) (ABTS),2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric reducing antioxidant power (FRAP) and protein oxidative damage protection assays. Compared with heating-only treated oat flour, that treated with α-amylase showed significant increase of extractable total phenolic content (0.46–1.35 μmol gallic acid equivalents per gram oat), total antioxidant capacity, and an increased ability on the protection of protein from oxidative damage. Heating-only increased caffeic acid and vanillin content by 17 (0.03 vs 0.54 μg/g oat) and 1.8 (0.62 vs 1.11 μg/g oat) folds, but slightly increased the content of other phenols. Excluding heating effect, α-amylase treatment increased gallic acid content by 2.6 folds (0.38 vs 1.38 μg/g oat), caffeic acid content by 2.4 (0.54 vs 1.82 μg/g oat) folds, and other phenols by 1.0–1.8 folds. In conclusion, α-amylase treatment can yield oat products containing more extractable phenolic compounds with increased antioxidant capacity.     

水杨酸诱导的水稻对白叶枯病的系统抗性与未处理叶酶活性的变化

 0.5、2.0 mmol/L 的水杨酸（SA）处理水稻幼苗后第3 天用稻白叶枯菌（Xanthomonas oryzae pv. oryzae）对未处理叶（第3叶，进行SA处理时用塑料袋套住）挑战接种，2周后调查病情，发现病斑长比对照分别下降了10.2%和18.2%，说明SA能诱导水稻幼苗对白叶枯病的系统抗性。酶活性测定表明，两种浓度的SA能明显降低水稻幼苗未处理叶中过氧化氢酶（CAT）和抗坏血酸过氧化物酶（APX）的活性，而对过氧化物酶（POD）活性无明显影响。两种浓度的SA对未处理叶中超氧化物歧化酶（SOD）活性的影响不同，其中0.5 mmol/L 的SA处理下，SOD无明显变化，但20 mmol/L 的SA对SOD活性有较明显的促进作用。SA处理后1～4 d，未处理叶H2O2含量比对照上升了23.6%～72.8％(0.5 mmol/L)和31.2%～122.6％(2.0 mmol/L)。H2O2水平的显著提高可能是H2O2产生加快和降解减慢的共同结果（2.0 mmol/L的SA下），或单纯的降解减慢所致（0.5 mmol/L的SA下）。同时，SA对未处理叶PAL活性有促进作用。提示在水稻中，SA对水稻抗白叶枯病的系统诱导作用可能与H2O2积累和PAL活性的升高有关。     

Inhibition of potato sprout growth by carvone enantiomers and their bioconversion in sprouts

Summary The monoterpenes (R)-(−)-carvone and (S)-(+)-carvone inhibited sprout growth in a model system consisting of sprouts growing from potato eye pieces. The sprout tissue was not necrotic after carvone treatment and the inhibition was reversible, since after treatment the sprouts showed regrowth either by continued top growth or by branching. However, the effect of both isomers on sprout growth differed, and (S)-(+)-carvone inhibited the elongation of the sprouts sooner than did (R)-(−)-carvone. This might be explained by a faster uptake of the former, since the concentration of (S)-(+)-carvone and its derivatives was twice as high during the first 4 days compared with (R)-(−)-carvone-treated sprouts. The sprouts were able to reduce (R)-(−)-carvone mainly into neodihydrocarveol, and (S)-(+)-carvone into neoisodihydrocarveol; in addition hydroxylated compounds were also detected. To whom correspondence should be addressed     

二斑叶螨为害前后抗、感木薯转录组分析及水杨酸、茉莉酸途径差异表达基因验证

本研究通过比较二斑叶螨为害前后抗、感螨木薯品种转录组差异,筛选差异表达基因,并采用qPCR验证水杨酸、茉莉酸信号途径基因的差异表达。转录组分析结果表明,与螨害前相比,螨害1、8 d后,抗螨木薯品种C1115的差异表达基因为589、587个,感螨木薯品种BRA900的差异表达基因为1271、930个,C1115相对于BRA900的差异表达基因分别为383、251个。GO和KEGG富集分析发现这些差异表达基因主要显著集中在次生代谢物质合成、苯丙烷生物合成、类黄酮生物合成和氧化还原反应等过程。qPCR验证结果显示,二斑叶螨为害后,抗螨参照标准木薯品种C1115的水杨酸信号途径基因PAL2、4CL3、WRKY7和NPR3的表达量较为害前呈现先显著提高后降低的趋势,而感螨参照标准木薯品种BRA900中上述4个基因的表达始终维持在显著高于为害前的水平。受螨害后抗螨木薯C1115中茉莉酸信号途径基因JAR1、LOX2和OPR11的表达量也较为害前显著提高,而感螨木薯BRA900中这3个基因的表达量则降低至显著低于螨害前的水平,qPCR验证结果和转录组分析结果相一致。本研究结果表明,抗螨木薯品种C1115受螨害后能够同时激活水杨酸、茉莉酸信号途径以抵御二斑叶螨为害,为深入阐明木薯抗螨分子机理,选育和创制抗螨木薯品种提供了理论参考依据。     

The effect of growth regulators on meristem tip development and in vitro multiplication ofSolanum tuberosum L. plants

Summary We studied the effect of β-indoleacetic acid (IAA), α-naphthylacetic acid (NAA), kinetin (K), 6-benzylaminopurine (BAP) and gibberellic acid (GA₃) on development of isolated meristems of four potato cultivars (subsp. tuberosum). GA₃ had a stimulating effect on in vitro meristem growth. The differences among the cultivars are presented. K+IAA+GA₃ induced formation of multiple shoots. We describe a method of continual plant multiplication under in vitro conditions and propose the produce for maintenance, multiplication and virus eradication of potato germplasm and cultivars.

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Zusammenfassung Von vier Kartoffelsorten (Blanik, Cira, Nora und Radka) wurden die Meristemspitzen entnommen. Der N?hrboden enthielt Makround Mikroelemente gem?ss Murashige & Skoog (1962), Vitamine entsprechend B-5 (Gamborg et al., 1968), 100mg/l Inositol, 30 g/l Saccharose und 8 g/l Agar. Wir untersuchten die Wirkung von β-Indolessigs?ure (IAA), α-Naphytlessigs?ure (NAA), Kinetin (K), 6-Benzylaminopurin (BAP) und Gibberellins?ure (GA₃). Auf den N?hrb?den mit IAA und NAA wurde mangelhaftes Triebwachstum und unvollst?ndige Blattentwicklung (Abb. 1) festgestellt; die Pflanzen konnten nicht in Erde versetzt werden. K (0.1 μM) induzierte vollst?ndige Pflanzenentwicklung (Tabelle 1), w?hrend BAP (0,1μM) die Triebenentwicklung anregte, worauf die Bewurzelung auf N?hrboden mit 0,1 μM IAA (Abb. 2) erfolgte. Alle GA₃-Konzentrationen hatten einen vorteilhaften Einfluss auf die Entwicklung von Pfl?nzchen aus Meristemspitzen (Abb. 1 und Tabelle 1). Optimales Wachstum und rasche Entwicklung der Pflanzen aller vier Sorten wurde mit dem N?hrboden mit IAA+K+GA₃ erreicht. Mehrfache Triebbildung erfolgte auf den Medien mit erh?hten GA₃-Konzentrationen (5 und 10 μM), speziell bei den Sorten Cira und Blanik (Abb. 3). Die Pflanzen aus der Meristemspitzenkultur konnten unter sterilen Bedingungen in Perlit, das mit der Hoagland's-L?sung und mit einem Zusatz von 0,1 μM IAA ges?ttigt war, fortlaufend vermehrt werden. Die Pflanzen entwickeln sich aus Achselknospen. Ein Steckling aus einem Meristem ist f?hig, 5–8 Pflanzen in zwei Monaten hervorzubringen. Der Artikel beschreibt das System der mehrfachen Triebbildung und die laufende Fortpflanzung in vitro zur Erhaltung und Vermehrung von Kartoffel-Keimplasma (Westcott et al., 1977) und virusfreien Züchtungen (Abb. 4).

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Résumé Des méristèmes apicaux ont été prélevés sur quatre variétés de pommes de terre: Blanik, Cira, Nora et Radka. Le milieu de culture contenait des macro-et microéléments d'après les indications de Murashige & Skoog (1962), des vitamines B-5 (d'après Gamborg et al., 1968), 100 mg/l d'inositol, 30 g/l de sucrose et 8 g/l d'agar. Nous avons étudié l'action de l'acide β-indoleacetic (IAA), de l'acide α-naphtylacétique (NAA), de la kinétine (K), de la 6-benzylaminopurine (BAP) et de l'acide gibberellic (GA₃). Sur les milieux contenant de l'IAA et du NAA, nous avons observé un développement imparfait des pousses, et un développement partiel des feuilles (fig. l); les plantes ne pouvaient pas être transplantées en terre. K (0,1 μM) a induit un développement complet des plantes (tableau 1) tandis que le BAP (0,1 μM) a stimulé le développement des pousses suivi de leur enracinement dans le milieu contenant 0,1 μM d'IAA (fig. 2). Toutes les concentrations de GA₃ ont eu un effet bénéfique sur le développement des plantules issues de méristèmes apicaux (fig. 1 et tableau 1). La croissance optimale et le développement rapide des plantes des 4 variétés ont été observés sur le milieu contenant IAA+K+GA₃. Des pousses multiples se sont formées sur les milieux contenant des concentrations élevées en GA₃ (5 et 10 μM), notamment sur les variétés Cira et Blanik (fig. 3). Les plantes obtenues à partir de culture de méristèmes apicaux ont p? être continuellement propagées en conditions stériles dans de la perlite saturée d'une solution de Hoagland avec addition de 0,1 μM de IAA. Les plantes se développent à partir des bourgeons axillaires; une plantule d'origine méristématique est capable de donner naissance à 5 à 8 plantes en 2 mois. L'article décrit le système de formation des pousses multiples et la propagation continue in vitro pour la maintenance et la multiplication de variétés de pommes de terre indemnes de maladies (Westcott et al., 1977) et de virus (fig. 4).

     

Characterizing the pigmented traditional rice cultivars grown in temperate regions of Kashmir (India) for free and bound phenolics compounds and in vitro antioxidant properties

The purpose of the research was to identify the phenolic and flavonoid compounds of seven different traditional pigmented whole rice cultivars grown in the temperate regions of Kashmir so as to study their relationship with in vitro antioxidant capacities. The completely pigmented rice cultivars were found to have higher phenolic, flavonoid, anthocyanin contents and exhibited higher antioxidant capacities than the light colored and sparely colored rice cultivars. A total of 40 compounds had been identified in the analyzed rice cultivars that were found to be distributed in 6 major categories with 6-phenolics, 6-flavonoids, 11-hydroxycinnamic acid derivatives, 7-hydroxybenzoic acid derivatives, 3-anthocyanins and 7-flavonoid glucosides of different flavonoid compounds. Among the free and bound fractions for each cultivars the light and sparsely colored depicted higher content of phenolics and in vitro antioxidant properties in bound faction, while the completely pigmented cultivars showed higher antioxidant properties in free fractions. The anthocyanins quercetin-3-O-galactoside, cyanidin-3-O-rutinoside and pelargonidin-3-O-diglucoside had been identified by LC-MS existing in the free fractions of the analyzed rice cultivars whereas, the free fraction of acetone + H₂O possessed higher percentage of phenolic compounds as compared to methanolic extracts and bound fractions. The black colored cultivars possessed higher DPPH scavenging activity and lipid peroxidation inhibition.     

Phytic acid,in vitro protein digestibility,dietary fiber,and minerals of pulses as influenced by processing methods

The objective of this project was to determine the effect of various types of processing on selected nutrition related parameters of commonly consumed Indian pulses and soybean. Germination reduced the phytic acid content of chickpea and pigeonpea seeds by over 60%, and that of mung bean, urd bean, and soybean by about 40%. Fermentation reduced phytic acid contents by 26–39% in all these legumes with the exception of pigeonpea in which it was reduced by more than 50%. Autoclaving and roasting were more effective in reducing phytic acid in chickpea and pigeonpea than in urd bean, mung bean, and soybean. Germination and fermentation greatly increased the in vitro protein digestibility (IVPD). IVPD was only slightly increased by roasting and autoclaving of all legumes. Germination and fermentation also remarkably decreased the total dietary fiber (TDF) in all legumes. Autoclaving and roasting resulted in slight increases in TDF values. All the processing treatments had little effect on calcium, magnesium and iron contents.     

Prevention of berry formation in potato plants (Solanum tuberosum L.) by single foliar applications of herbicides or growth regulators

Summary Thre seed production in potato plants (Solanum tuberosum L.) may contribute to the persistence of potato cyst nematodes and of viruses, fungi and bacteria. In four experiments MCPA, ethephon, 2,4D-amine, naphtylacetamide, metoxuron and gibberellic acid were tested for their potential to reduce berry formation. MCPA (500 or 750 g a.i. ha⁻¹ and ethephon (144 or 192 g a.i. ha⁻¹) reduced or almost completely prevented berry set and their effect was strongly influenced by the crop growth stage at the time of application. For both chemicals application at early bud stage was the most effective and, whereas MCPA showed no adverse effects on tuber yield, size, dry matter content and fry colour, ethephon substantially reduced the percentage of large tubers over 50 mm.     

Improvement of growth and gymnemic acid production by altering the macro elements concentration and nitrogen source supply in cell suspension cultures of Gymnema sylvestre R. Br

Gymnema sylvestre is an important medicinal plant which bears bioactive compound namely gymnemic acids. The present work deals with optimization of cell suspension culture system of G. sylvestre for the production of biomass and gymnemic acid and we investigated effects of macro elements (NH₄NO₃, KNO₃, CaCl₂, MgSO₄ and KH₂PO₄ - 0.0, 0.5, 1.0, 1.5 and 2.0× strength) and nitrogen source [NH₄⁺/NO₃⁻ ratio of: 0.00/18.80, 7.19/18.80, 14.38/18.80, 21.57/18.80, 28.75/18.80, 14.38/0.00, 14.38/9.40, 14.38/18.80, 14.38/28.20 and 14.38/37.60 (mM)] of Murashige and Skoog medium on accumulation of biomass and gymnemic acid content. The highest accumulation of biomass (165.00 g l⁻¹ FW and 15.42 g l⁻¹ DW) was recorded in the medium with 0.5× concentration of NH₄NO₃ and the highest production of gymnemic acid content was recorded in the medium with 2.0× KH₂PO₄ (11.32 mg g⁻¹ DW). The NH₄⁺/NO₃⁻ ratio also influenced cell growth and gymnemic acid production; both parameters were greater when the NO₃⁻ concentration was higher than that of NH₄⁺. Maximum biomass growth (159.72 g l⁻¹ of FW and 14.95 g l⁻¹ of DW) was achieved at an NH₄⁺/NO₃⁻ ratio of 7.19/18.80, and gymnemic acid production was also greatest at the same concentration of NH₄⁺/NO₃⁻ ratio (11.35 mg g⁻¹ DW).