植物春化作用的分子机制及应用

植物能响应环境中的低温信号实现开花调控，这种经历低温促使植物开花的作用即为春化作用。本文结合国内外春化作用的研究进展，概述春化作用的发现和特点，阐述模式植物拟南芥及单子叶作物的春化作用分子机制，并举例说明春化作用在农业、园艺等方面的应用。     

冬小麦春化过程中可溶性蛋白质组成的变化与形态发生的关系

冬小麦“农大139”经40天左右的春化处理才能迅速而整齐地抽穗,但经14—21天低温处理,已经具有在夏季抽穗的可能性,虽然抽穗推迟且极不整齐;再将春化时间延长,则抽穗百分比增加,且从播种到抽穗的时间缩短。这表明,春化过程中低温对发育的作用有两种效应:前期低温是诱发生理状态的转变,后期低温则只具有加速发育的作用,两个时期的转变是在春化的中期。蛋白质合成抑制剂乙基硫氨酸和对-氟苯丙氨酸能抑制冬小麦的春化,抑制时期也是在春化过程的中期。不同时间低温处理后冬小麦幼芽中可溶性蛋白质含量及组成发生了变化,春化过程中期(低温处理14天之后)不仅含量比对照增加了一倍,而且有新的蛋白质谱带出现。春麦中无类似现象,未经低温处理的春麦已含有冬麦中新出现的谱带。说明冬小麦春化过程的第14—21天左右是与春化过程有关的蛋白质合成的关键时期,该时期新合成的蛋白质与植株的发育状态之间存在着密切的相关关系。     

冬小麦春化作用相关cDNA克隆Vrc79的分子克隆

在低温需求型植物的发育过程中,春化作用是诱导植物成花的必要因子。在冬小麦春化进程中存在着一个核酸代谢的关键期,为了探讨春化作用的分子机理,以不春化及脱春化的冬小麦京冬1号（Ｔｒｉｔｉｃｕｍ　ａｅｓｔｉｖｕｍＬ．ｃｖ．Ｊｉｎｄｏｎｇ　Ｎｏ．１）胚芽的ｍＲＮＡ为对照,通过减法杂交构建了富集冬小麦春化相关基因的ｃＤＮＡ文库,经过差异筛选分离到了一个仅在春化２１ｄ这一关键期表达,而在不春化、春化４ｄ、脱春化不表达的春化相关。ｃＤＮＡ克隆Ｖｒｃ７９。Ｎｏｒｔｈｅｍ杂交及同源性分析表明它是一个在植物中新发现的不同于低温胁迫诱导基因的春化特异克隆,其对春化需求型植物的成花诱导可能起重要的调控作用。     

植物春化特性及春化作用机理

春化作用是某些高等植物成花转变的重要环节,被认为是植物在低温诱导下促使其相关基因的表达,从而导致生理状态转变的一种受遗传控制的生理过程。本文对植物春化反应特性、春化作用的生理生化特性及春化作用的分子生物学等方面的研究进展进行了概述,并对春化研究中的问题进行了分析和展望。     

植物春化特性及春化作用机理

春化作用是某些高等植物成花转变的重要环节,被认为是植物在低温诱导下促使其相关基因的表达,从而导致生理状态转变的一种受遗传控制的生理过程.本文对植物春化反应特性、春化作用的生理生化特性及春化作用的分子生物学等方面的研究进展进行了概述,并对春化研究中的问题进行了分析和展望.     

植物如何消除春化的记忆——ELF6阻止春化状态遗传给下一代

<正>植物能够感知周围环境,控制和协调自身的发育,使其在合适的时间开花结果。春化过程就是一个很好的例子,植物通过感受冬天连续的低温环境,逐渐抑制开花抑制因子的表达,并且能够在冬季之后保持该基因的抑制状态,即植物通过表观修饰可以在随后的发育过程中记住基因的表达状态,从而促进开花。春化对于许多冬性作物(如小麦、大麦)的高产、稳产也具有重要作用。模式植物拟南芥和作物通过不同的开花抑制因子(例如:拟南芥中FLC和小麦及大麦中VRN2)响应春化过程,但研究认为它们具有类似的调节机制。认识拟南芥中FLC的调节对     

冬小麦春化相关基因cDNA(verc203)片段序列的分析

春化作用是高等植物发育的重要环节之一,它是受遗传控制的生理过程,人们对此现象已作了生态、生理和生化方面的研究（谭克辉1983）。认为植物茎尖生长点接受低温诱导后,会引起体内许多代谢途径和方式的顺序改变（种康和谭克辉1993）,可能导致春化相关基因启动表达。有人（Burn等1993）提出春化相关基因启动表达可能是基因脱甲基化结果的观点。但这一假说尚缺乏直接的强有力证据。违斌和谭克辉（199）系统地研究了冬小麦春化过程中核酸、蛋白质代谢与春化作用的直接相关关系和春化特异蛋白质代谢与特异性rnRNA出现的重要意义,确信春化…     

植物非编码RNA调控春化作用的表观遗传

在自然界中许多高等植物需要通过冬季的低温阶段实现从营养生长到生殖生长的时期转化,这一生物学过程称作春化作用。小麦(Triticum aestivum L.)和油菜(Brassica napus L.)等作物以种子为产品器官,生产上往往通过茬口安排和栽培措施使植株尽早通过春化作用,以促进花芽形成和花器官发育,而大白菜(B rapa ssp.pekinenesis)和甘蓝(B.oleracea)等作物以叶球等营养器官作为产品器官,生产上则设法避免低温引起的春化作用,以保证产品器官的充分生长。FLOWERING LOCUS C(FLC)作为一种重要的开花抑制蛋白负调控春化作用,参与植株从营养生长向生殖生长的转化过程。文章综述了春化中FLC表达受抑制主要通过低温诱导表达FLC基因区域的非编码RNA以及VRN1、VRN2、VIN3等蛋白参与介导组蛋白甲基化,从而在表观遗传上控制春化作用的进程和产品器官的正常发育。     

高等植物春化作用研究进展

环境因子(低温诱导或称春化作用)在高等植物的生长发育调控中起着重要作用。本文就近年来春化作用诱导的开花启动研究领域中的进展从春化过程中的多步骤性、成花决定过程控制的多途径性、接受春化刺激的位置与分化启动的关系、以及春化作用的分子机理等几个方面进行了简要综述。并对春化相关基因克隆方面的进展进行了分析和综述。     

“先驱”转录因子LEC1在早期胚胎重置春化状态的机制

开花是植物由营养生长阶段向生殖生长阶段转变的重要过程,长时间低温处理即春化对开花起到非常重要的促进作用。春化控制的拟南芥(Arabidopsis thaliana)开花中,阻抑型转录因子FLC是重要的关节点,春化记忆依赖于对该基因的控制。何跃辉研究组之前对拟南芥的研究揭示了转录因子VAL1或VAL2可以识别负调控开花的关键基因FLC成核区的顺式DNA元件,协同PRC2复合体在春化过程中沉默FLC基因的表达,并在随后的常温下继续维持FLC基因沉默直至受精结束,使植物产生春化记忆。但在下一代中如何擦除这种记忆功能,使FLC重新被激活,以防止植物在过冬前或过冬时开花,相关机制目前并不清楚。近期,该研究组揭示了在植物胚胎发育早期一个种子特有的"先驱"转录因子参与擦除春化记忆,重新激活FLC基因的分子机制,并解析了胚胎中的基因激活传递到后胚胎发育(营养生长期)的表观遗传机理。该研究是开花领域的重要突破,为作物开花调控的生产应用提供了新思路。     

Zearalenone-Like Substance in Winter Plants and Its Relation to Vernalization

It was found that there is a zearalenone-like substance in the growing points of vernalized wheat seedlings and carrot plants. The content of this substance synchronously increased with the depth of vernalization. It was separated by thin layer chromatography. Its UV spectrum was similar to that of zearalenone. It is believed that this substance may be one of the important endogenous substances which control the vernalization of winter plants.     

Efficient shoot and root formation from cotton shoot apices

Successful shoot and root induction were obtained from shoot apices of two cotton (Gossypium hirsutum L.) genotypes, Nazilli 84S and Çukurova 1518, which are widely planted in Turkey. Plant tissue culture systems were established on Murashige and Skoog (MS) medium supplemented with various plant growth regulators using seven-day-old shoot apices as explants. The shoot apex size was of 2–3 mm; it contained the meristem and unexpanded leaves. Shoot apices were placed on MS plus vitamins and combinations of various plant hormones. The best regeneration responses were obtained for cv. Nazilli 84S (98%) on MS + 0.1 mg/l kinetin (KIN) + 1 g/l polyvinylpyrrolidone (PVP) and for Çukurova 1518 (94%) on MS + 0.1 mg/l KIN + 2 mg/l NAA + 1 g/l PVP. Including germination, all regeneration and rooting processes lasted only 5 weeks. The shoot apices of both genotypes developed successfully without intervening callus formation, and no significant differences between cultivars were found. All regenerated plants of both genotypes were phenotypically normal and set seeds. This shoot meristem-based rapid regeneration method can also be used in the cases of biolistic and Agrobacterium-mediated transformation.     

In vitro clonal multiplication of an apple rootstock by culture of shoot apices and axillary buds

In vitro clonal multiplication of apple rootstock MM 111 using axillary buds and shoot apices were carried out. Vegetative axillary buds of the size of 0.2-2.0 cm and shoot apices measuring 4 mm in length were initiated to shoot proliferation on MS medium supplemented with BA (0.5 - 1.0 mgl(-1)), GA3(0.5 mgl(-1)), with or without IBA(0.05 - 0.1 mgl(-1)). Small size explants showed less phenol exudation and less contamination. Following establishment phase, the small shoots emerged from explants were subcultured on MS medium supplemented with different combinations and concentrations of growth regulators. BA (1.0 mgl(-1)) and GA3 (0.5 mgl(-1)) combination showed highest multiplication rate (1:5), andcl also produced longer shoots. Two step rooting was done by transferring microcuttings to auxin free solid medium after root initiation in dark on 1/2 strength MS liquid medium containing IBA (0.5 mgl(-1) ). Rooted plantlets were transferred to peat containing paper cups and resulting plants of MM 111 acclimated successfully for transfer to field.     

Appearance of peroxidase isozymes in floral‐initiated shoot apices of Pharbitis nil

A novel biochemical assay system for detecting the early stage of flowering is reported. Peroxidase isozymes in shoot apices of Pharbitis nil plants that had been exposed to flower‐inducing or non‐inducing conditions were analyzed by isoelectric focusing in polyacrylamide gels and activity staining for peroxidase. Several isozymes with pH 8.5–8.8 appeared for the first time 7 days after the beginning of short‐day treatment, but not after nightbreak (non‐inducing) treatment. When shoot tips were cultured in vitro, these same isozymes also appeared after short‐day treatment but not after night‐break treatment. The extent of the appearance of these isozymes was reduced by exposure to high or low temperature during the inductive dark period and removal of cotyledons after the inductive dark period. Such treatments also reduced the extent of flowering. The appearance of an isozyme with pH 8.5 was more closely correlated with flowering than that of the other isozymes. From these results, the appearance of this peroxidase isozyme in shoot apices is discussed as a biochemical marker of flowering in intact plants and in cultured shoot tips.     

Restricted cell/cell communication in the shoot apex ofSilene coeli-rosa during the transition to flowering is associated with a high mitotic index rather than with evocation

Summary Plants ofSilene coeli-rosa given 5 or more long days (LDs) flowered, even when the LDs were followed by 48 hours of darkness before their return to short days (SDs). The mitotic indices of shoot apices from induced plants shortly after induction were significantly higher than the indices of shoot apices from vegetative plants. Two major mitotic peaks were observed in the shoot apices of plants given 7 long days (LDs) on day 8. One coincided with that reported byFrancis andLyndon (1979).Cell to cell movement was tested in the shoot apices of vegetative and LD treated plants using probes with a molecular size of 749 daltons (fluorescein-hexaglycine) and 847 daltons (fluorescein-leucyl diglutamyl leucine). These probes showed some movement in the shoot apices of both short day (SD) and LD treated plants, but fluorescein-leucyl diglutamyl leucine was immobile in the induced apices of 7 LD plants on day 8 at time intervals which coincided with major mitotic activity in the shoot apex. Symplasmic restriction in the shoot apex was also observed in plants given 8 LDs (i.e., plants not returned to SDs on day 7).In plants that were placed in 48 hours of darkness after the 7 LD treatment or in plants given 5 LDs, there was no strong peak in the mitotic index, even though all these LD treatments resulted in 100% flowering. In such plants no symplasmic restriction was found in the shoot. Thus the symplasmic restriction on day 8 of 7 LD plants is associated with the high mitotic index, but neither of these phenomena is an essential part of the evocation process.Abbreviations F(Glu)₂ L-glutamylglutamic acid conjugated to fluorescein isothiocyanate isomer I (F-) - F(Gly)₆ F-hexaglycine - FLGGL F-leucyl-diglutamyl-leucine - F(PPG)₅ F-the pentamer (propyl-propyl glycine) - LD long day - LDs long days - SD short day - SDs short days     

STRUCTURAL AND HISTOLOGICAL STUDIES OF THE CAMBIUM AND SHOOT MERISTEMS OF SOYBEAN TREATED WITH 2,3,5-TRIIODOBENZOIC ACID

Flowering soybeans were sprayed at the tips with 50 ppm TIBA. Microscopic and macroscopic observations were made of the nodes, internodes, and shoot meristems every week for 4 wks after TIBA treatment. TIBA-treated plants produced open flowers at the upper nodes 1 week earlier than did control plants. Accompanying this early flower development, the following changes occurred in the upper internodes, as compared to controls: (a) increased activity of the procambium; (b) rapid development of thick-walled protophloem cells; (c) production of small vessels. Three weeks after treatment middle internodes of treated plants showed less cambial activity than did corresponding internodes of controls. The changes in the middle internodes of treated plants suggest a close correlation with increased flowering. Two weeks after treatment some lateral shoot apices at nodes nearest the main shoot apex exhibited the following changes, in contrast with controls: (a) development of conical apices with stack-of-brick-like peripheral cells; (b) shrinkage of protoplasmic contents in some rib meristem cells and young pith cells; (c) frequent thickening of primary walls in young pith cells. Three weeks after treatment cells of lateral shoot meristems with conical axillary buds showed a denser stain for protein than did cells of corresponding meristems of controls. Floral apices in these meristems also stained more densely for protein than did similar apices in control plants. Together with early flower production in the upper nodes of treated plants, less starch occurred 2–3 weeks after treatment than in corresponding nodes of controls. Two to three weeks after treatment lateral shoot apices of both treated and control plants had numerous, large starch grains in the rib meristem, young pith, leaf and bud primordia, and developing flowers but few starch grains appeared in the tunica, corpus, and procambium.     

Amelioration of salinity tolerance in <Emphasis Type="Italic">Solanum tuberosum</Emphasis> L. by exogenous application of ascorbic acid

The present investigation envisaged revealing the role of exogenous application of ascorbic acid in increasing resistance against NaCl stress. Shoot apices from 60-d-old, in vitro-grown plants of two commercially important cultivars of Solanum tuberosum L., cvs. Desiree and Cardinal, were inoculated on Murashige and Skoog (MS) medium supplemented with 0.5 mM ascorbic acid for 72 h as a pretreatment. Pretreated and non-pretreated shoot apices were transferred to MS medium containing different concentrations of NaCl (0–140 mM; eight treatments). Results were recorded for morphological (shoot length, shoot number, root length, root number, and number of nodes) and biochemical features (protein, peroxidase, catalase, and superoxide dismutase activities) after 60 d of salt treatment. Similarly, 60-d-old, well-proliferated callus cultures were also pretreated with ascorbic acid for 24 h and transferred to an optimized callus proliferation medium containing different concentrations of salt. Results were recorded after 60 d of salt treatment for percentage relative fresh weight growth and biochemical parameters. Salinity severely inhibited all the growth parameters in both the cultivars. Pretreatment with ascorbic acid to both salt-treated plants and callus cultures showed significant differences with respect to almost all of the growth and biochemical parameters studied. Protein content as well as catalase and superoxide dismutase activities increased significantly in both the cultivars, although peroxidase activity showed a decreasing trend in ascorbic acid-pretreated plants as well as callus cultures.     

THE EFFECTS ON CHRONIC GAMMA RADIATION ON THE INTERNAL APICAL CONFIGURATIONS OF THE VEGETATIVE SHOOT APEX OF COLEUS BLUMEI

Clonal plants of a bushy variety ‘Trailing Queen’ of Coleus blumei Benth. were exposed to 600–700 R per day for 28 days. Shoot apices of control and irradiated plants were studied microscopically. The normal vegetative shoot apex has two tunica layers and a corpus which is partly layered and partly composed of randomly divided cells. Five cytohistological zones were observed, including a cup-shaped zone, imposed upon the tunica-corpus relationships. Response to gamma radiation resulted in abnormal internal apical configurations. All irradiated shoots was found to have the same internal configurations. Only a single tunica layer broken by occasional periclinal divisions was observed. The cytohistological organization of irradiated apices was different from that seen in the normal shoot tips in that two zones were found. Evidence of a radiosensitivity gradient within the vegetative shoot apex was also observed. The possible significance of the results is discussed.