植物染色体G—显带的AEG法

目前,植物染色体G—显带的方法很多,有胰酶法、尿素法,改良的Seabright法、Utakoji法、醋酸地衣红法、ASG法和风油精诱导等几种,都取得了显著的效果。在广泛实验研究的基础上,我们摸索出了     

植物染色体G—带的研究进展

染色体 G-带技术在动物和人类遗传学中已得到了广泛应用,可是在植物方面由于它仅能显示带纹很少的 C-带、N-带或 Q-节,这就大大限制了染色体显带技术在植物细胞遗传学研究和植物育种上的进一步应用。近十年来,植物染色体 G-带的研究越来越受到人们的重视,世界上,尤其是我国有不少学者进行了详细研究,并取得了不少进展,本文仅就这方面的研究现状做一简述。     

植物染色体G带研究概况

本文对植物染色体G带的历史、认识发展、技术探索、现状、问题及前景等进行了全面综述。显带技术建立后的多年,植物染色体一直不显G带,不少研究者提出了不同假说予以解释或进行实验验证。最近几年,该技术有了迅速发展,尤其是我国学者在这方面做了大量开创性工作,用多种方法在十几种植物上进行了探索,并初获成功。虽然目前还存在一些有待解决的困难和问题,但植物染色体G带技术有着广阔的发展前景。     

应用G显色带染色体荧光原位杂交（FISH）技术研究复杂的染色体易位

建立常规Ｇ显带染色体标的荧光原位杂交（ＦＩＳＨ）技术,用于分析患者复杂的染色体易位,原位杂交前,用甲醛固定Ｇ显带标本,是获得良好显带和荧光杂交效果的关键步聚,仅用常细胞遗学方法分析,显示一例习惯性流产患者的核型为４６,ＸＸ,ｔ（１;５;１２）（１ｐｔｅｒ→１ｑ２５：：１２ｑ２４→１２ｑｔｅｒ;５ｑｔｅｒ→５ｑ１１：：１ｑ２５→１ｑｔｅｒ;１２ｐｔｅｒ→１２ｑ２４;：５ｐ１１→５ｐｔｅｒ）而采用本方     

水稻染色体G—带的研究

用改良的ASG法首次在籼稻(O.sativa subsp.indica)品种珍汕97和粳稻(O.subsp.iaponica)品种秀岭的有丝分裂染色体上显示了G-带,并作了相应的G-带核型分析。就同一材料来说,随着有丝分裂时期的推进,染色体上带纹数目逐渐减少。籼、粳亚种间相对应的同源染色体上G-带带纹特征彼此相似。讨论了水稻G-带带型与染色体不同区域分化的关系;G-带带型与籼、粳稻分歧的关系;以及G-显带的方法。     

应用G显带染色体荧光原位杂交（FISH）技术研究复杂的染色体易位

建立常规Ｇ显带染色体标本的荧光原位杂交（ＦＩＳＨ）技术,用于分析患者复杂的染色体易位。原位杂交前,用甲醛固定Ｇ显带标本,是获得良好显带和荧光杂交效果的关键步骤。仅用常规细胞遗传学方法分析,显示一例习惯性流产患者的核型为４６,ＸＸ,ｔ（１;５;１２）（１ｐｔｅｒ→１ｑ２５：：１２ｑ２４→１２ｑｔｅｒ;５ｑｔｅｒ→５ｐ１１：：１ｑ２５→１ｑｔｅｒ,１２ｐｔｅｒ→１２ｑ２４：．５ｐ１１→５ｐｔｅｒ）,而采用本方法确定患者的核型实际为４６,ＸＸ,ｔ（１;５,１２）（１ｐｔｅｒ→１ｑ２３：：１２ｑ２２→１２ｑｔｅｒ,５ｑｔｅｒ→５ｐ１１：：１ｑ２５→１ｑｔｅｒ;１２ｐｔｅｒ→１２ｑ２２：：１ｑ２３→１ｑ２５：５ｐ１１→５ｐｔｅｒ）。结果表明,新建立的Ｇ显带染色体荧光原位杂交（ＦＩＳＨ）技术能更有效地检测患者复杂的染色体易位。     

鱼类染色体G—显带的Brd U—BSG方法及白鲢G—带模式图的初步建立

本文报道了一种显示鱼类染色体G-带的BrdU-BsG方法。采用肾细胞短期培养,收获前12小时加入BrdU,使终浓度为10μg/ml。制片经HCl、Ba(OH)_2处理,4×SSC温育。Giemsa染色,显示出白鲢的G-带。其带纹细致清晰,一个细胞的单倍染色体上显示带纹达200条以上,是目前已报道的鱼类多重带中带纹最多的,且反差明显,带纹有特征性,结果较稳定。根据实验结果初步建立了白鲢的G-带模式图。     

玉米染色体G—带带型的研究

本文对3个玉米自交系,及其中两个自交系的杂交F_1有丝分裂早中期染色体的G-带带型进行了比较研究。所有的供试材料G-显带的染色体上都具有两种类型的带纹,我们称A型带和B型带。A型带为沿染色体长轴分布,较细的,密切邻近的多重带纹。不同自交系的A型带带型基本相同,杂交F_1的A型带无明显的异型性。非同源染色体间带型各不相同,某些染色体具有易于识别,特征性较强的A型带标记。B型带一般为深染色的大带,位于染色体的近端区。同一自交系每两个同源染色体的B型带可以配对,不同自交系B型带带型互有不同。杂交F_1某些染色体上的B型带带型异型性明显。具异型性的染色体对中一成员的带型与一个亲本相似,另一成员与另一亲本相似。比较对同一细胞先后作G-和C-显带处理的结果表明,B型带和C-带是相同的。     

Presence of molybdenum cofactor in seeds of wheat and barley

Molybdenum cofactor (Mo-co) was determined in seeds of wheat and barley by its ability to restore nitrate reductase (NR) activity in extracts of nitrate reductase-deficient mutants. Its activity was compared with that of wheat roots and leaves. Conditions for assay of Mo-co from different sources in the presence of molybdate and reduced glutathione (GSH) were optimised. The effect of heat-treatment of cell-free extracts from seeds, roots and leaves was also investigated. Mutant extracts of Neurospora crassa nit-1 and Nicotiana tabacum CnxA68, used as apoprotein source for in vitro complementation, were shown to give comparable results. The Mo-co activity, extracted from wheat and barley seeds, could be separated into two peaks by gel chromatography.     

Abscisic acid content and growth of spring barley plants treated with triazoles

Effects of triazolium compounds (paclobutrazol, uniconazole, and azovit), applied under laboratory and greenhouse conditions, on growth of barley plants (Hordeum vulgare L., cvs. Moskovskii 2 and Zazerskii 85) and ABA content in their tissues were studied for the first time. At various application techniques, the retardants suppressed growth of axial organs, promoted thickening and strengthening of lower internodes, and caused a considerable increase in ABA content. These changes were observed already on the next day after spraying plants with retardants and maintained for two weeks after the spraying treatment. At the same time, the retardant action sustained for 42 days when the retardants were applied to soil. The triazoles had positive effects on plant productivity characteristics, such as the number of filled grains in the main spike, total weight of grain per plant (grain collected from the main and lateral spikes), and the weight of 1000 grains. It was concluded that the retardant effect of triazoles on spring barley plants is related to elevated ABA content in plant tissues over a prolonged growth period.Translated from Fiziologiya Rastenii, Vol. 52, No. 1, 2005, pp. 108–114.Original Russian Text Copyright © 2005 by Chizhova, Pavlova, Prusakova.     

A relationship between humidity response, growth form and photosynthetic operating point in C3 plants

Gas exchange experiments were performed with 13 plant species that differ from each other in growth-form and natural habitat. These comprised three herbaceous species, two ferns, two temperate deciduous trees, five rainforest trees and one liana from wet tropical forest. The aims were to investigate whether plants of similar growth-form and from similar habitats tended to respond similarly to a change in leaf-to-air vapour pressure difference (VPD), and to compare their ratio of intercellular to ambient partial pressures of CO₂ for given conditions. Leaves were subjected to a step change in VPD and the initial and final steady rates of transpiration were used to calculate an index of sensitivity, φ , which enabled comparison of species. The results suggest that species of similar growth-form and habitat respond similarly to increasing VPD, with the temperate deciduous trees undergoing a greater reduction in stomatal conductance than the herbaceous plants in well-watered soil. Also, for these experimental conditions, the ratio of leaf internal to ambient CO₂ partial pressure (p_i/p_a) was positively correlated with both CO₂ assimilation rate and stomatal insensitivity to VPD, across the 13 species. The results are discussed in terms of growth strategies and possible advantages and limitations of hydraulic systems in different plants.     

Waterlogging tolerance in the tribe Triticeae: the adventitious roots of Critesion marinum have a relatively high porosity and a barrier to radial oxygen loss

Nine species from the tribe Triticeae – three crop, three pasture and three ‘wild’ wetland species – were evaluated for tolerance to growth in stagnant deoxygenated nutrient solution and also for traits that enhance longitudinal O₂ movement within the roots. Critesion marinum (syn. Hordeum marinum) was the only species evaluated that had a strong barrier to radial O₂ loss (ROL) in the basal regions of its adventitious roots. Barriers to ROL have previously been documented in roots of several wetland species, although not in any close relatives of dryland crop species. Moreover, the porosity in adventitious roots of C. marinum was relatively high: 14% and 25% in plants grown in aerated and stagnant solutions, respectively. The porosity of C. marinum roots in the aerated solution was 1·8–5·4‐fold greater, and in the stagnant solution 1·2–2·8‐fold greater, than in the eight other species when grown under the same conditions. These traits presumably contributed to C. marinum having a 1·4–3 times greater adventitious root length than the other species when grown in deoxygenated stagnant nutrient solution or in waterlogged soil. The length of the adventitious roots and ROL profiles of C. marinum grown in waterlogged soil were comparable to those of the extremely waterlogging‐tolerant species Echinochloa crus‐galli L. (P. Beauv.). The superior tolerance of C. marinum, as compared to Hordeum vulgare (the closest cultivated relative), was confirmed in pots of soil waterlogged for 21 d; H. vulgare suffered severe reductions in shoot and adventitious root dry mass (81% and 67%, respectively), whereas C. marinum shoot mass was only reduced by 38% and adventitious root mass was not affected.     

Distribution of free and bound forms of cis-trans and trans-trans abscisic acid in broad-bean plants in relation to apical dominance

Levels of endogenous abscisic acid (ABA; free and bound forms) have been determined by gas chromatography in stems and buds of broad-bean plants ( Vicia faba L. cv. Aguadulce) in relation to apical dominance. A downward gradient of free cis-trans ABA occurred along the stem, from the apical bud to the roots. Except for the actively growing apical bud the levels of free cis-trans ABA were higher in the buds than in the corresponding nodes. An inverse correlation can be set up between levels of free cis-trans ABA and growth of buds, except for the cotyledonary ones. High levels of bound ABA ( cis-trans form) are correlated with the growth of the apical bud and that of the axillary bud ax1. The hormonal regulation of the growth of the cotyledonary buds, which contained high levels of trans-trans ABA in bound forms, is apparently different from that of the other buds.     

Distribution of labelled auxin and derivatives in stem tissues of intact and decapitated broad-bean plants in relation to apical dominance

[³H]-auxin (0.13 to 0.18 nmol) was applied to the apical bud of broadbean plants (Vicia faba L. cv. Aguadulce). After 24 h, the exportation from the donor organ was ended. After 48 h, i.e. 10–15 h after the passage of the [³H]-auxin pulse into the root system, the distribution and the nature of labelled molecules located in the basal part of the stem and in the axillary buds were investigated. Chromatographic analyses concerned both intact plants and plants decapitated 12 h, 24 h or 42 h after the [³H]-auxin application. In intact plants, there was no significant amount of [³H]-auxin in the axillary buds, whose radioactivity was very low compared to the stem tissues. The labelled molecules with the Rf of auxin represented 50% or more of the whole radioactivity of the stem tissues. The distribution of [³H]-auxin was not uniform along the stem. In particular, the cotyledonary node zone, bearing the most inhibited buds, which is known to be an important centre of label retention, contained the highest amounts of labelled auxin both in intact and decapitated plants. The decapitation was quickly followed by a decrease of the [³H]-auxin amount in the stem base more than 15 cm away from the wound, particularly in the scale leaf nodes, whose axillary buds were mainly the ones to grow after relief from apical dominance. The induction of this early decrease was clearly distinct in plants decapitated when auxin exportation from the donor organ was ended.     

Drought- and ABA-induced changes in photosynthesis of barley plants

The changes caused by drought stress and abscisic acid (ABA) on photosynthesis of barley plants (Hordeum vulgare. L. cv. Alfa) have been studied. Drought stress was induced by allowing the leaves to lose 12% of their fresh weight. Cycloheximide (CHI), an inhibitor of stress-induced ABA accumulation, was used to distinguish alterations in photosynthetic reactions that are induced after drought stress in response to elevated ABA levels from those that are caused directly by altered water relations. Four hoars after imposition of drought stress or 2 h after application of ABA, Ihe bulk of the leaf's ABA content measured by enzyme-amplified ELISA, increased 14- and 16-fold, respectively. CHI fully blocked the stress-induced ABA accumulation. Gas exchange measurements and analysis of enzyme activities were used to study the reactions of photosynthesis to drought stress and ABA. Leaf dehydration or ABA treatment led to a noticeable decrease in both the initial slope of the curves representing net photosynthetic rate versus intercellular CO₂ concentration and the maximal rate of photosynthesis; dehydration of CHI-treated plants showed much slower inhibition of the latter. The calculated values of the intercellular CO₂ concentration, CO₂ compensation point and maximal carboxylating efficiency of ribulose 1,5-bisphosphate (RuBP) carboxylase support the suggestion that biochemical factors are involved in the response of photosynthesis to ABA and drought stress. RuBP carboxylase activity was almost unaffected in ABA- and CHI-treated, non-stressed plants. A drop in enzyme activity was observed after leaf dehydration of the control and ABA-treated plants. When barley plants were supplied with ABA, the activity of carbonic anhydrase (CA, EC 4.2.2.1) increased more than 2-fold. Subsequent dehydration caused an over 1.5-fold increase in CA activity of the control plants and a more than 2.5-fold increase in ABA-treated plants. Dehydration of CHI-treated plants caused no change in enzyme activity. It is suggested that increased activity of CA is a photosynthetic response to elevated ABA concentration.     

Comparative studies of diurnal variations of nitrate reductase activity in wheat, oat and barley

Diurnal variations of in vitro and in vivo (intact tissue assay) nitrate reductase (EC 1.6.6.1) activity and stability were examined in leaves of wheat ( Triticum aestivum L. cv. Runar), oat ( Avcna saliva L. cv. Mustang) and barley ( Hordeum vulgure L. cv. Agneta and cv. Gunillu). Nitrate reductase activity was generally higher for wheat than for oat and barley. However, the diurnal variations of nitrate reductase activity and stability were principally the same for all species, e.g. the high activity during the photoperiod was associated with low stability. All species showed a rapid (30-60 min) increase in the in vitro and in vivo activity when the light was switched on. When light was switched off the in vitro activity decreased rapidly whereas decrease in in vivo activity was slower. These experiments support the hypothesis that an activation/ deactivation mechanism is involved in the regulation of diurnal variations in nitrate reductase activity. Red light enhanced nitrate reductase activity in etiolated wheat and barley leaves. In green leaves, however, the daily increase in nitrate reductase activity was not induced by a brief red light treatment. Indications of different regulation mechanisms for the diurnal variations of nitrate reductase activity among the cereals were not found.     

Development and accumulation of ABA in fluridone-treated and drought-stressed Vicia faba plants under different light conditions

The effects of fluridone on guard cell morphology, chloroplast ultrastructure and accumulation of drought stress-induced abscisic acid (ABA) were studied in Vicia faba L. plants grown under different light conditions. Drought stress was induced by allowing the leaves to lose 12% of their fresh weight. The appearance of defective and undeveloped stomata, and chloroplasts with a destroyed thylakoid membrane system was found in fluridone-treated plants grown at a photosynthetic photon flux (PPF) of 600 μmol m^-2 s^-1. Plants grown at a PPF of 40 μmol m^-2 s^-1 had diminished levels of ABA after imposition of dehydration. Fluridone treatment reduced the level of ABA in both unstressed and dehydrated leaves. Accumulation of ABA in the control plants was considerably reduced when they were exposed to dark periods of 24, 48 and 72 h just before imposition of the stress. Twenty-four hours after the dark treatment dehydration of the leaves resulted in a 3-fold decrease in the level of stress-induced ABA, and 72 h after dark treatment the amount of stress-induced ABA approximated the prestressed values. Fluridone-treated plants failed to accumulate ABA under water stress. In addition to functionally active chloroplasts, well-developed and functional stomata are required for drought stress to elicit a rise in ABA.