Effect of polyamines on adventitious root formation from Tobacco (Nicotiana tabaccum) leaf segments

To elucidate the effect of polyamines on adventitious root formation, we investigated the relationship between the frequency of adventitious root formation and the endogenous content of free polyamines in tobacco leaf segments which had been treated with polyamine biosynthesis inhibitors and polyamines. Adventitious root formation was inhibited in rooting medium (10 μM IAA) with methylglyoxal-bis(guanylhydrazone) (MGBG) or cyclohexylamine (CHA), and promoted with spermidine and putrescine. Treatment with high IAA (100 μM) medium plus CHA or MGBG promoted rooting up reversion of the rooting inhibition than the one treated with high IAA concentration alone. Spermidine promoted adventitious root numbers on low IAA (1 μM) medium when applied during culture period. The rooting inductive phase (in the presence of IAA) was determined by periodical transfer of leaf segments from IAA-containing medium to IAA free medium, and by changing polyamine contents, to be inductive phase. Putrescine and spermidine were accumulated to a maximum during the inductive phase. Therefore, the results point out the involvement of polyamines in inductive phase of adventitious root formation in tobacco leaf segments.     

Chloroplast chlB gene is required for light-independent chlorophyll accumulation in Chlamydomonas reinhardtii

Light-independent chlorophyll synthesis occurs in some algae, lower plants, and gymnosperms, but not in angiosperms. We have identified a new chloroplast gene, chlB, that is required for the light-independent accumulation of chlorophyll in the green alga Chlamydomonas reinhardtii. The chlB gene was cloned, sequenced, and then disrupted by performing particle gun-mediated chloroplast transformation. The resulting homoplasmic mutant was unable to accumulate chlorophyll in the dark and thus exhibited a yellow-in-the-dark phenotype. The chlB gene encodes a polypeptide of 688 amino acid residues, and is distinct from two previously characterized chloroplast genes (chlN and chlL) also required for light-independent chlorophyll accumulation in C. reinhardtii. Three unidentified open reading frames in chloroplast genomes of liverwort, black pine, and Chlamydomonas moewusii were also identified as chlB genes, based on their striking sequence similarities to the C. reinhardtii chlB gene. A chlB-like gene is absent in chloroplast genomes of tobacco and rice, consistent with the lack of light-independent chlorophyll synthesis in these plants. Polypeptides encoded by the chloroplast chlB genes also show significant sequence similarities with the bchB gene product of Rhodobacter capsulatus. Comparisons among the chloroplast chlB and the bacterial bchB gene products revealed five highly conserved sequence areas that are interspersed by four stretches of highly variable and probably insertional sequences.     

Carbon monoxide promotes root hair development in tomato

This study identified the role of CO in regulating the tomato root hair development. Exogenous CO promoted the root hair density and elongation in a concentration-dependent manner. Analysis of cross sections of primary roots also indicated that CO induced the formation of root hairs. Genetic analysis reveals that tomato mutant yg-2 (defective in haem oxygenase-1 activity and intracellular CO generation) displayed a phenotype of delayed root hair development, which however could be reversed by exogenous CO. Further, we analysed LeExt1 :: β -glucuronidase reporter gene for root hair formation and found increasing expression of LeExt1 in the CO-exposed root hairs. Finally, CO was able to act synergistically with auxin, ethylene and NO. It is shown that the effect of CO could be blocked by NPA (auxin transport inhibitor), AVG (ethylene biosynthesis inhibitor), Ag⁺ (ethylene action inhibitor) or cPTIO (NO scavenger). Exposure of tomato roots to CO also enhanced intracellular NO and reactive oxygen species generation in root hairs. Our results suggest that CO would be required for root hair development and may play a critical role in controlling architectural development of plant roots by a putative mechanism of cross-talk with auxin, ethylene and nitric oxide.     

Endophytic actinomycetes from Pinus thunbergii and their antifungal activity against Cylindrocladium sp.

The inside of Pinus thunbergii could be a reliable screening source for a useful agent in controlling plant disease. Isolation of endophytic actinomycetes from P. thunbergii and their potential as biocontrol agents against the plant pathogen Cylindrocladium sp. were investigated. Two endophytic actinomycetes, Streptomyces sp. and Microbispora sp., were isolated from surface-sterilised root tissues of P. thunbergii seedlings. The recovery test of these two endophytic actinomycetes from pine seedling showed that Streptomyces sp. was isolated from only roots, but Microbispora sp. was isolated from both roots and leaves. Thus, Microbispora sp. is able to move to leaves from roots. Moreover, we evaluated the potential of both strains as biocontrol agents against Cylindrocladium sp. Two weeks after inoculation of Cylindrocladium sp. alone, pine seedlings showed a 50% mortality rate. Co-inoculation of Cylindrocladium sp. and Microbispora sp. did not affect seedling mortality rate. However, inoculation with both Cylindrocladium sp. and Streptomyces sp. reduced seedling mortality to 12%. Streptomyces sp. could be a useful agent in controlling pine disease caused by Cylindrocladium sp. Thus, it seems that Streptomyces sp. may induce a local host defence reaction and Microbispora sp. systemically spreads to aerial parts through the transpiration stream.     

烟草主要数量性状的遗传效应分析

利用红花大金元×青梗,红花大金元×中烟14号P1、P2、F1、F2、B1、B2 6个世代资料对7个农艺性状和4个品质性状进行了基因效应分析。结果表明,性状均不符合简单的加性－显性遗传模型,多数性状加性效应显著而显性效应不显著,在3种互作效应中,所有性状至少有一种显著。互作效应普遍存在,是烟草性状杂种优势表现的主要原因之一。 Abstract:Two tobacco F1 hybrids,F2s,backcrosses B1s and B2s and their parents P1 and P2 were used to estimate the gene effects for 7 agronomic and 4 quality characters.The additive-dominance genetic model was not fit for all characters.The additive effects and the epistatic effects of most characters were significant,but the dominant effect not.The epistatic effects could not be ignored in tobacco breeding.They were one of main causes of heterosis for most characters.     

马尾松雌球果的发生和早期发育研究

采用常规石蜡制片技术对马尾松雌球果的发生和早期发育进行了研究。结果表明：雌球果原基发生时间为10月中旬,不同的树龄和着生部位,其发生时间不同。雌球果原基与营养茎端在外部形态及内部细胞组织学分区结构有明显差异。营养茎端外形扁平,内部顶端分生组织结构有顶端原始细胞区、中央母细胞区、形成层状过渡区、周围分生组织区及肋状分生组织区5个明显的分区;而雌球果原基外形呈圆锥状,内部结构只有套层和髓区。12月初,最初的苞片原基在雌球果原基的鳞片的叶腋处产生,之后其由基部向顶部连续发生。翌年1月初,在苞片原基的叶腋处,珠鳞原基发生,发生方向亦为向顶发育。2月底,苞片体积不再发生变化,珠鳞膨大端的基部的近轴面分化出2个倒生胚珠。从雌球果原基发生到胚珠分化历时4个多月。亚热带的冬季气候对马尾松雌球果的生长发育没有明显的抑制作用。     

Product enhancement and recovery from transformed root cultures of Nicotiana glauca

Transformed roots of Nicotiana glauce synthesize the alkaloids nicotine and anabasine at levels reflecting the parent plants. Media composition, strength, and pH were evaluated with respect to biomass yield and productivity. Full-strength Gamborg's B5 medium proved the best for biomass yield while half-strength, or low-salt, medium enhanced alkaloid accumulation. A detailed investigation of media nitrate levels demonstrated how these may be manipulated to promote growth and intracellular or extracellular alkaloid levels. High nitrate concentrations were found to significantly enhance media alkaloid levels at the end of the growth phase. Media pH is also important, although transformed roots will grow in Gamborg's B5 medium between pH 3 and 9, root biomass is favored by an increase in medium alkalinity, while alkaloid release is encouraged by mildly acidic pH.Transformed roots release a proportion of their secondary metabolites into the growth medium. By continually removing root products, any feedback inhibition on enzymatic reactions is reduced, as are the toxic effects resulting from product accumulation. In this article we describe the use of Amberlite resins (XAD-2 and XAD-4) to enhance alkaloid levels (nicotine and anabasine) of hairy root cultures of Nicotiana glauca by a factor of 10 with no adverse effect on root growth. The performance of the Amberlite columns was subsequently investigated with respect to alkaloid adsorption and desorption, including an evaluation of the effects of pH and loading capacity. The resins also adsorb media constituents which are identified and quantified as part of this work. Resulting nutritional stresses are thought to be partly responsible for enhancing secondary metabolism at the expense of biomass yield. However, the net effects of using Amberlite resins as a means of product removal significantly increases the overall product yield and the extent to which products are released into the growth medium.     

沙质海岸黑松分枝格局特征及其抗风折能力分析

沙质海岸空间梯度上环境差异较大, 黑松(Pinus thunbergii)在长期的适应过程中树冠结构变化也很大。为揭示黑松树冠结构与环境间的适应机制, 在山东省胶南市灵山湾国家森林公园距海岸线0-50、200-250和400-450 m梯度内各设置1个样带, 记为带I、带II、带III, 采用枝构型的理论和方法, 对黑松的分枝格局进行了研究, 并采用模拟自然风法测定了黑松枝条的抗风折能力。结果表明: 1)在海岸梯度上黑松分枝格局差异较大, 随着距离海岸越来越远, 黑松各级枝的分枝长度、总体分枝率均逐渐增大, 而枝径比和逐步分枝率逐渐减小, 各级分枝角度表现为带I >带III >带II。2)带I树冠背风面与迎风面相比, 分枝长度、分枝角度、分枝数量、枝条干枯率分别是迎风面的1.62、1.38、2.65和0.59倍, 随着距离海岸越来越远, 这种不对称性逐渐减弱, 至带III树冠基本对称。3)海风是影响带I分枝角度偏转、枝条干枯和冠型不对称现象的主要原因。4)带III枝条的抗风折能力高于带I, 且两个样带模拟风速与枝条所承受的拉力之间的关系均符合逻辑斯蒂方程, 相关系数R²均达0.97以上。该研究揭示了不同海岸梯度上黑松分枝格局的形成机制及其抗风折能力, 可为沿海黑松防护林的合理经营提供科学依据。     

黑松与松材线虫互作早期NO应答信号与外在因子的关系

松树与松材线虫互作中,NO作为重要的信号分子参与调控寄主对松材线虫入侵的应答反应,但松材线虫入侵寄主后,是松材线虫还是其代谢产物激发寄主体内NO应答信号尚不明确.本研究用松材线虫活体、线虫分泌物、虫体研磨液等分别接种黑松,研究接种早期黑松体内NO合成酶(NOS)活性和NO含量变化与后期松树症状发展的关系.结果表明:采用虫体研磨液、线虫分泌物处理后黑松体内NOS活性和NO含量均升高;各处理黑松外部均出现感病症状.说明在松树与松材线虫互作中,除线虫活体作用外,线虫体或分泌物中的物质也可激发松树体内NO应答信号的表达,诱导下游应答响应的发生,导致黑松感病.15 ～25℃条件下,随着温度升高,接种黑松体内NOS活性增强,NO含量升高,感病症状出现较早;随着干旱胁迫程度的增强,接种黑松体内NOS活性、NO含量大致呈逐步升高趋势,感病症状也较早出现.在一定范围内,温度升高和干旱程度加剧可诱发感病黑松体内NO相关信号途径表达增强,并加速病程发展.表明在一定条件下,高温干旱环境有利于松材线虫病的发生可能与寄主体内NO应答信号增强有关.     

海岸梯度上黑松针叶形态与解剖结构性状的变化规律

风对树木形态、生理和生长的影响一直是生态学研究的热点和难点问题,目前主要采用模拟风速或机械刺激的方式来研究树木对风胁迫的响应与适应变化,无法准确反映出树木在风环境下的长期适应机制。本文以海岸60年生黑松林（Pinus thunbergii）为对象,分析了海岸距离梯度上黑松迎风面和背风面针叶长度、宽度、面积、比叶面积、角质层厚度、表皮厚度等形态和解剖结构特征的变化规律。结果表明：黑松迎风面针叶长度、宽度、周长、面积、厚度和比叶面积随着离海岸距离的减小逐渐减小（P<0.05）;而表皮厚度、角质层厚度和导管孔径则随着离海岸距离的减小逐渐增加（P<0.05）;背风面各指标均无明显变化规律。黑松迎风面与背风面针叶性状（比叶面积除外）主要在海岸500 m内存在显著差异（P<0.05）。可以看出,随着海风胁迫的增加,黑松通过降低针叶长、宽等形态指标,有效减少了风胁迫下黑松的受力面积,是一种躲避策略;而增加针叶表皮、角质层厚度和导管孔径等解剖结构指标,则增强了针叶坚韧性和保证针叶的水分供应,是一种忍受策略,这些策略有利于该树种在大风环境中得以更好的生存。     

Coordinating early kidney development: lessons from gene targeting

The kidney is widely used to study the mechanisms of organogenesis. Its development involves fundamental processes, such as epithelial branching, induced morphogenesis and cytodifferentiation, which are common to the development of many other organs. Gene-targeting experiments have greatly improved our understanding of kidney development, and have revealed many important genes that regulate early kidney organogenesis, some of which have a role in inherited human kidney disorders. Although our understanding of how the kidney is assembled is still limited, these studies are beginning to provide insights into the genetic and cellular interactions that regulate early organogenesis.     

Young Leaf Chlorosis 1, a chloroplast-localized gene required for chlorophyll and lutein accumulation during early leaf development in rice

Chlorophyll (Chl) and lutein are the two most abundant and essential components in photosynthetic apparatus, and play critical roles in plant development. In this study, we characterized a rice mutant named young leaf chlorosis 1 (ylc1) from a ⁶⁰Co-irradiated population. Young leaves of the ylc1 mutant showed decreased levels of Chl and lutein compared to those of wild type, and transmission electron microscopy analysis revealed that the thylakoid lamellar structures were obviously loosely arranged. Whereas, the mutant turns green gradually and approaches normal green at the maximum tillering stage. The Young Leaf Chlorosis 1 (YLC1) gene was isolated via map-based cloning and identified to encode a protein of unknown function belonging to the DUF3353 superfamily. Complementation and RNA-interference tests confirmed the role of the YLC1 gene, which expressed in all tested rice tissues, especially in the leaves. Real-time PCR analyses showed that the expression levels of the genes associated with Chl biosynthesis and photosynthesis were affected in ylc1 mutant at different temperatures. In rice protoplasts, the YLC1 protein displayed a typical chloroplast location pattern. The N-terminal 50 amino acid residues were confirmed to be necessary and sufficient for chloroplast targeting. These data suggested that the YLC1 protein may be involved in Chl and lutein accumulation and chloroplast development at early leaf development in rice.     

美国黄松组织培养不定根诱导的研究

以GD、SH和1／2SH基本培养基对美国黄松不定芽进行不定根的诱导。试验结果表明基本培养基的种类对不定芽形成不定根起主要作用。在1／2SH培养基上附加0．5mg／L的NAA不定根的诱导率为3．3％。试验首次在离体培养条件下,以美国黄松种胚为外植体获得了再生小植株。     

Abscisic acid promotes pre-emergence stages of lateral root development in Medicago truncatula

The plant root system is important for plant anchorage and nutrition. Among the different characteristics of the root system, root branching is a major factor of plasticity and adaptation to changing environments. Indeed, many biotic and abiotic stresses, such as drought or symbiotic interactions, influence root branching. Many studies concerning root development and root branching were performed on the model plant Arabidopsis thaliana, but this model plant has a very simplified root structure and is not able to establish any symbiotic interactions. We have recently described 7 stages for lateral root development in the model legume Medicago truncatula and found significant differences in the tissular contribution of root cell layers to the formation of new lateral roots (LR). We have also described 2 transgenic lines expressing the DR5:GUS and DR5:VENUS-N7 reporter genes that are useful to follow LR formation at early developmental stages. Here, we describe the use of these transgenic lines to monitor LR developmental responses of M. truncatula to the phytohormone abscisic acid (ABA) which is a major actor of stress and symbiotic interactions. We show that ABA promotes the formation of new lateral root primordia and their development, mostly at the late, pre-emergence stage.