Expansin-regulated cell elongation is involved in the drought tolerance in wheat

Water stress restrains plant growth. Expansin is a cell wall protein that is generally accepted to be the key regulator of cell wall extension during plant growth. In this study, we used two different wheat cultivars to study the involvement of expansin in drought tolerance. Wheat coleoptile was used as the material in experiment. Our results indicated that water stress induced an increase in acidic pH-dependant cell wall extension, which is related to expansin activity; however, water stress inhibited coleoptile elongation growth. The increased expansin activity was mainly due to increased expression of expansin protein that was upregulated by water stress, but water stress also resulted in a decrease in cell wall acidity, a negative factor for cell wall extension. Decreased plasma membrane H⁺-ATPase activity was involved in the alkalinization of the cell wall under water stress. The activity of expansin in HF9703 (a drought-tolerant wheat cultivar) was always higher than that in 921842 (a drought-sensitive wheat cultivar) under both normal and water stress conditions, which may be correlated with the higher expansin protein expression and plasma membrane H⁺-ATPase activity observed in HF9703 versus 921842. However, water stress did not change the susceptibility of the wheat cell wall to expansin, and no difference in this susceptibility was observed between the drought-tolerant and drought-sensitive wheat cultivars. These results suggest the involvement of expansin in cell elongation and the drought resistance of wheat.     

Effect of ABA on the water relations of winter-wheat cultivars varying in drought resistance

Water and osmotic potentials were measured in leaves of a drought-sensitive ('Ponca') and a drought-resistant ('KanKing') cultivar of winter wheat ( Triticum aestivum L . em. Thell.) to determine if the potentials of the drought-sensitive cultivar could be made similar to those of the drought-resistant cultivar through application of abscisic acid (ABA). Stomatal resistance was also measured. Plants were sprayed with ABA and grown in soil, which was watered or allowed to dry. In well-watered plants, ABA closed the stomata of both cultivars. Stomatal resistance of plants grown without added water and with ABA was less than that of plants grown without added water and without ABA. Under ample water supply, ABA decreased water and osmotic potentials of the drought-sensitive cultivar (Ponca), but had no effect on these potentials in the drought-resistant cultivar (KanKing). Under water-deprived conditions, ABA increased water and osmotic potentials of Ponca, but did not change these potentials in KanKing. The overall effect of ABA was to decrease the differences in the water and osmotic potentials between the two cultivars.     

Expansins and coleoptile elongation in wheat

Expansins are now generally accepted to be the key regulators of wall extension during plant growth. The aim of this study was to characterize expansins in wheat coleoptiles and determine their roles in regulating cell growth. Endogenous and reconstituted wall extension activities of wheat coleoptiles were measured. The identification of beta-expansins was confirmed on the basis of expansin activity, immunoblot analysis, and beta-expansin inhibition. Expansin activities of wheat coleoptiles were shown to be sensitive to pH and a number of exogenously applied factors, and their optimum pH range was found to be 4.0 to 4.5, close to that of alpha-expansins. They were induced by dithiothreitol, K(+), and Mg(2+), but inhibited by Zn(2+), Cu(2+), Al(3+), and Ca(2+), similar to those found in cucumber hypocotyls. An expansin antibody raised against TaEXPB23, a vegetative expansin of the beta-expansin family, greatly inhibited acid-induced extension of native wheat coleoptiles and only one protein band was recognized in Western blot experiments, suggesting that beta-expansins are the main members affecting cell wall extension of wheat coleoptiles. The growth of wheat coleoptiles was closely related to the activity and expression of expansins. In conclusion, our results suggest the presence of expansins in wheat coleoptiles, and it is possible that most of them are members of the beta-expansin family, but are not group 1 grass pollen allergens. The growth of wheat coleoptiles is intimately correlated with expansin expression, in particularly that of beta-expansins.     

Role of Expansin in Cell Enlargement of Oat Coleoptiles (Analysis of Developmental Gradients and Photocontrol)

Expansins are wall proteins that mediate a type of acid-induced extension in isolated plant cell walls (S. McQueen-Mason, D.M. Durachko, D.J. Cosgrove [1992] Plant Cell 4: 1425-1433). To assess the role of these proteins in the process of cell enlargement in living tissues, we compared the spatial and temporal growth patterns of oat (Avena sativa L.) coleoptiles with four wall properties related to expansin action. These properties were (a) the ability of isolated walls and living segments to extend in acidic buffer, (b) the ability of heat-inactivated walls to extend upon application of expansins, (c) the amount of immunologically detectable expansin in wall protein extracts, and (d) the extractable expansin activity of walls. Growth rate was maximal in the apical half of dark-grown coleoptiles and negligible in the basal region. This growth pattern correlated with properties a and b; in contrast, the amount and activity of extractable expansin (properties c and d) were reduced only in the most basal region. Upon exposure to white light, coleoptiles abruptly ceased elongation at 8 to 10 h after start of irradiation, and this cessation correlated with reductions in properties a to c. The growth cessation at 8 to 10 h also coincided with the loss of growth response to exogenous auxin and fusicoccin in excised coleoptile segments. These results lend correlative support to the hypothesis that expansin action is important for growth responses of living oat coleoptiles (e.g. responses to acidic buffers, auxin, fusicoccin, aging, and light). Our results suggest that changes in the susceptibility of the wall to expansin action, rather than changes in expansin activity, may be a key determinant of the growth patterns in oat coleoptiles.     

Inhibition of the Coleoptile Growth in Avena sativa by Endosperm Removal--Changes in Auxin, Osmotica and Cell Wall

Removal of the endosperm from 84-h-old etiolated oat seedlingsstrongly retarded the subsequent growth of coleoptiles. Thecontribution of the endosperm to coleoptile growth was studied.Endosperm removal was found to: (1) decrease the endogenouslevel of indole-3-acetic acid (IAA) in the coleoptile tip. IAAapplied to the coleoptile tip stimulated coleoptile growth inseedlings with and without the endosperm. The sensitivity ofthe coleoptile to a suboptimal concentration of IAA was higherin seedlings without the endosperm than in intact ones. At theoptimal concentration of IAA, however, the final length of thecoleoptile was larger in intact seedlings than in those withoutthe endosperm. (2) decrease the concentration of the solublesugars and amino acids in the cell sap. (3) retard the increasein the amount of polysaccharides in the cell wall of the coleoptile,particularly noncellulosic ones. (4) make the cell wall mechanicallyrigid according to stress-relaxation analysis of the cell wall.(5) induce an increase in the osmotic potential of the coleoptilecell sap. From these results, it was concluded that the endosperm suppliesthe coleoptile with IAA, sugars and amino acids, thus promotingcoleoptile growth. (Received September 24, 1987; Accepted February 3, 1988)     

Inhibition of the synthesis of cell wall polysaccharides in oat coleoptile segments by jasmonic acid: Relevance to its growth inhibition

The inhibitory mode of action of jasmonic acid (JA) on the growth of etiolated oat (Avena sativa L. cv. Victory) coleoptile segments was studied in relation to the synthesis of cell wall polysaccharides using [¹⁴C]glucose. Exogenously applied JA significantly inhibited indoleacetic acid (IAA)-induced elongation of oat coleoptile segments and prevented the increase of the total amounts of cell wall polysaccharides in both the noncellulosic and cellulosic fractions during coleoptile growth. JA had no effect on neutral sugar compositions of hemicellulosic polysaccharides but substantially inhibited the IAA-stimulated incorporation of [¹⁴C]glucose into noncellulosic and cellulosic polysaccharides. JA-induced inhibition of growth was completely prevented by pretreating segments with 30 mm sucrose for 4 h before the addition of IAA. The endogenous levels of UDP-sugars, which are key intermediates for the synthesis of cell wall polysaccharides, were not reduced significantly by JA. Although these observations suggest that the inhibitory mode of action of JA associated with the growth of oat coleoptile segments is relevant to sugar metabolism during cell wall polysaccharide synthesis, the precise site of inhibition remains to be investigated.Abbreviations JA jasmonic acid - ABA abscisic acid - IAA indoleacetic acid - T ₀ minimum stress relaxation time - TFA trifluoroacetic acid - TCA trichloroacetic acid - HPLC high-performance liquid chromatography - EtOAc ethyl acetate - TLC thin-layer chromatography - JA-Me methyl jasmonate - GLC-SIM gas-liquid chromatography-selected ion monitoring     

扩展蛋白与植物抗逆性关系研究进展

扩展蛋白是一种细胞壁蛋白,可调节细胞壁的松弛和伸展。目前研究表明,扩展蛋白几乎参与调节植物生长发育的整个进程。扩展蛋白还与植物的多种抗性反应有关,在植物对干旱、高盐以及病虫害等生物胁迫和非生物胁迫响应方面起着重要的调节作用。干旱胁迫下扩展蛋白基因的表达与植物的抗旱性有一定的关系;植物的耐盐性受到扩展蛋白基因表达的影响;淹水促进植物的伸长生长与扩展蛋白的表达密切相关;扩展蛋白调节细胞壁松弛为植物抗病性研究提供了新的思路。     

RADIOAUTOGRAPHIC STUDY OF CELL WALL DEPOSITION IN GROWING PLANT CELLS

Segments cut from growing oat coleoptiles and pea stems were fed glucose-³H in presence and absence of the growth hormone indoleacetic acid (IAA). By means of electron microscope radioautography it was demonstrated that new cell wall material is deposited both at the wall surface (apposition) and within the preexisting wall structure (internally). Quantitative profiles for the distribution of incorporation with position through the thickness of the wall were obtained for the thick outer wall of epidermal cells. With both oat coleoptile and pea stem epidermal outer walls, it was found that a larger proportion of the newly synthesized wall material appeared to become incorporated within the wall in the presence of IAA. Extraction experiments on coleoptile tissue showed that activity that had been incorporated into the cell wall interior represented noncellulosic constituents, mainly hemicelluloses, whereas cellulose was deposited largely or entirely by apposition. It seems possible that internal incorporation of hemicelluloses plays a role in the cell wall expansion process that is involved in cell growth.     

渗透胁迫下小麦根系渗透调节与根冠淀粉水解的研究

用不同浓度的PEG—600对抗旱性不同的小麦幼苗进行渗透胁迫处理,研究了小麦幼苗根系的淀粉酶活性、可溶性糖含量、渗透势、渗透调节能力和根冠淀粉的水解状况。结果表明,随着渗透胁迫程度的加重,抗旱性强的小麦品种昌乐5号和北农2号根系渗透势和饱和渗透势的降低程度大于抗旱性弱的小麦品种鲁麦5号和921842,并且抗旱性强的小麦品种根系的渗透调节能力大于抗旱性弱的小麦品种。随着渗透胁迫程度的加重．各品种小麦根冠淀粉粒均有不同程度的减少。而抗旱性强的品种根冠淀粉粒的减少程度小于抗旱性弱的品种;抗旱性强的小麦品种根系淀粉酶活性显著高于抗旱性弱的小麦品种,但是,随着渗透胁迫程度的加重,抗旱性弱的品种淀粉酶活性增加的幅度远高于抗旱性强的品种。可溶性糖含量的变化趋势与淀粉酶活性的变化趋势一致．即渗透胁迫下根冠淀粉水解程度大的小麦品种,可溶性糖的含量高。但根冠淀粉水解在根系的渗透调节以及在小麦适应水分胁迫中的作用还有待于进一步探讨。     

渗透胁迫对小麦胚芽鞘内多胺的种类、形态和含量的影响

用高压液相色谱法研究了豫麦18（抗旱性较强）和扬麦9号（抗旱性较弱）小麦胚芽鞘中三种不同形态的多胺（polyamine,PA）：游离态多胺（PA）、高氯酸可溶性结合态多胺（ps结合态PA）和高氯酸不溶性结合态多胺（PIS结合态PA）与渗透胁迫的关系。结果发现：渗透胁迫2d,豫麦18胚芽鞘中的游离态Spd和游离态Spm的含量明显上升,而扬麦9号的游离态Put的上升明显。S-腺苷蛋氨酸脱羧酶（S—AMDC）的抑制剂——甲基乙二醛-双（鸟嘌呤腙）（MGBG）处理豫麦18,明显抑制了渗透胁迫诱导的游离态Spd和游离态Spm的增加,并且加重了渗透胁迫伤害,外源Spd处理扬麦9号明显促进了渗透胁迫诱导的游离态Spd和游离态Spm的增加,并且减缓了渗透胁迫的伤害。渗透胁迫下,豫麦18胚芽鞘中的PS结合态PA和PIS结合态PA的上升幅度都明显大于扬麦9号。菲咯啉（o—Phen）抑制渗透胁迫下PIS结合态PA的合成并加重了渗透胁迫对胚芽鞘的伤害。这些结果表明：小麦胚芽鞘中的游离态Spd、游离态Spm、PS结合态PA和PIS结合态PA的升高有利于增强渗透胁迫抗性。     

Relationship between osmotic stress-induced abscisic acid accumulation,biomass production and plant growth in drought-tolerant and -sensitive wheat cultivars

The effects of increasing osmotic stress induced by 100–400 mOsm (−0.976 MPa) polyethylene glycol (PEG 6000) were investigated in a drought-tolerant (Triticum aestivum L. cv. Mv Emese) and drought-sensitive (cv. GK élet) wheat cultivar at the three-leaf stage. During osmotic stress, the decline of the water potential (ψ _w) was more significant in the leaves, while the abscisic acid (ABA) levels of the roots increased earlier and remained higher in the sensitive than in the tolerant variety. There was an increasing gradient of ABA content toward the youngest leaves in the drought-sensitive GK élet, while more ABA accumulated in the fully developed, older leaves of the tolerant cultivar Mv Emese. In accordance with the rapid and significant accumulation of ABA, the stomatal conductance decreased earlier in the tolerant cultivar. The effect of water stress on the PSII photochemistry was pronounced only 1 week after the exposure to PEG, as indicated by the earlier decrease of the net CO₂ fixation, the effective quantum yield (Φ_PSII) and the photochemical quenching (q _P) in light-adapted samples of the tolerant variety in 400 mOsm PEG 6000. The stress treatment caused more significant reductions in these parameters toward the end of the experiment in the sensitive cultivar. In spite of small differences in the photosynthetic characteristics, the net biomass production was not significantly altered by this osmotic stress. The accumulation of ABA controlled the distribution of the biomass between the shoot and root systems under osmotic stress, and contributed to the development of stronger and deeper roots in the drought-sensitive cultivar GK élet. However, the root elongation did not correlate with the drought sensitivity of these cultivars on the basis of crop yield.     

The Geoelectric Effect in Plant Shoots:: IV. INTER-RELATIONSHIP BETWEEN GROWTH, AUXIN CONCENTRATION AND ELECTRICAL POTENTIALS IN ZEA COLEOPTILES

Incubation of Zea coleoptiles in 0.5 M mannitol totally inhibitsgrowth and geotropic curvature, but does not affect the developmentof the geoelectric effect. This pre-treatment also inhibitsthe curvature induced by the asymmetrical application of IAAto the apical end of decapitated vertical coleoptiles, but itdoes not prevent the IAA from giving rise to an electropotentialdifference between the two sides of the coleoptile. Neitherthe normal geoelectric effect, nor the auxin-induced potentialdifference in vertical coleoptiles, can therefore arise as theresult of the different rates of cell extension in the two halvesof the organ. They must be the result of the change of IAA concentrationaffecting some other aspect of the cell's physiology or metabolism. The abolition of the electrical responses in coleoptiles whichhave been plasmolysed in 1.0 M mannitol strongly suggests thatboth longitudinal and lateral transport of IAA are severelydepressed by this degree of plasmolysis. Asymmetrical application of 10^-5 M mersalyl and several othersubstances to the apical end of a decapitated vertical coleoptilegave rise to a marked electropotential difference between thetwo sides of the coleoptile, the side beneath the donor beingpositively charged with respect to the other side. Mersalyldoes not promote the growth of Zea coleoptiles. These resultsprovide additional evidence that the electropotentials do notarise from differential growth, and suggest that such substances,especially the diuretics used in clinical medicine, may provideuseful tools in the further study of the induction of surfaceelectropotentials in plant tissues at the cellular level.     

水分胁迫对抗旱性不同的荔枝叶片细胞壁H+-ATPase活性的影响

以抗旱性较强的荔枝东刘1号和抗旱性较弱的陈紫1－2年分盆栽实生幼苗为试验材料。研究了水分胁迫对荔枝（Litchi chinensis Sonn）叶片细胞胞质以及以离子键和共价键与细胞壁结合的H^ -ATPase活性的影响。结果表明：（1）叶片相对含水量随水分胁迫程度的增加而减少,抗旱性强的东刘1号下降的幅度小于抗旱性弱的陈紫。（2）H^ -ATPase在细胞中的分布是：细胞胞质H^ -ATPase占绝大多数,其次是共价键结合H^ -ATPase,离子键结合H^ -ATPase最少,品种间差异不明显。（3）在水分胁迫下,荔枝叶片H^ -ATPase活性（比活性）均上升,抗旱性强的品种上升的幅度均大于抗旱性弱的品种。     

Changes in cellular osmotic potential and mechanical properties of cell walls during light-induced inhibition of cell elongation in maize coleoptiles

The growth rate of maize ( Zea mays L. cv. Cross Bantam T51) coleoptiles in the dark was highest at the basal zone and decreased towards the tip. Growth was strongly inhibited by white fluorescent light (5 W m⁻²), especially in the basal zone of coleoptiles. Light irradiation caused an increase in the values of stress-relaxation parameters, the minimum stress-relaxation time and the relaxation rate and a decrease in the extensibility (strain/stress) of the cell walls at all zones. In addition, during growth, the accumulation of osmotic solutes was strongly inhibited by white light irradiation, resulting in an increased osmotic potential. The influences of white light on the mechanical properties of the cell wall and the osmotic potential of the tissue sap were most prominent in the basal zone. Significant correlations were observed between the increment of coleoptile length and the mechanical properties of the cell walls or the osmotic potential of the tissue sap and osmotic solutes content. Furthermore, light inhibited the outward bending of split coleoptile segments. These facts suggest that white light inhibits elongation of maize coleoptiles by modifying both the mechanical properties of the cell walls and cellular osmotic potential, which control the rate of water uptake.     

Osmoregulation in Rice Coleoptile Elongation as Promoted by Cooperation between IAA and Ethylene

IAA-induced elongation of rice (Oryza sativa L. cv. Sasanishiki)coleoptiles is regulated by cooperation between IAA and ethyleneproduced in response to IAA. However, the presence of some solutes,such as K^$, Na^$, Rb^$, glucose and sucrose, in the incubationmedia was found to be indispensable for this cooperation. Withoutthose solutes, the IAA-induced elongation was not sustainedover a long time period. IAA caused increases in both the osmoticpotentials of the coleoptile cells and the extensibility oftheir cell wall. In epidermal cells of IAA-treated coleoptiles,the osmotic potential increased from –0.87 to –0.62MPa during a 4-h incubation with 1 mM KCl. Moreover, IAA promotedthe uptake of K^$ or Na^$ from the media into the coleoptiles.However, these effects of IAA were partially prevented by aminoethoxyvinylglycine(AVG), and all the AVG effects were completely nullified byethylene applied simultaneously and exogenously. Both IAA andethylene did not affect the wall yield stress. These resultssuggested that the long-term elongation induced by IAA in ricecoleoptile segments results from inhibiting increases in osmoticpotentials of their cells. The maintenance by IAA of low osmoticpotentials may be partly due to the promotive action of ethyleneproduced in response to IAA on the solute uptake from the media. (Received July 6, 1983; Accepted February 15, 1984)     

Auxin-induced elongation growth and expressions of cell wall-bound exo- and endo-beta-glucanases in barley coleoptiles

When auxin stimulates rapid cell elongation growth of cereal coleoptiles, it causes a degradation of 1,3:1,4-beta-glucan in hemicellulosic polysaccharides. We examined gene expressions of endo-1,3:1,4-beta-glucanase (EI) and exo-beta-glucanase (ExoII), of which optimum pH are about 5, and molecular distribution of hemicellulosic polysaccharides in barley (Hordeum vulgare L.) coleoptile segments treated with or without IAA. IAA (10(-5) M) stimulated the gene expression of EI, while it did not affect that of ExoII. IAA induced gene expression of EI after 4 h and increased wall-bound glucanase activity after 8 h. The molecular weight distribution of hemicellulosic polysaccharides from coleoptile cell walls was shifted to lower molecular weight region by 2 h of IAA treatment. Fusicoccin (10(-6) M) mimicked IAA-induced elongation growth and the decrease in molecular weight of hemicellulosic 1,3:1,4-beta-glucan of coleoptiles in the first 4 h, but it did not promote elongation growth thereafter. These facts suggest that acidification of barley cell walls by IAA action enhances pre-existing cell wall-bound glucanase activity in the early first phase of IAA-induced growth and the late second phase involves the gene expression of EI by IAA.     

Differential changes in cell wall matrix polysaccharides and glycoside-hydrolyzing enzymes in developing wheat seedlings differing in drought tolerance

The growth kinetics and variations in cell wall matrix polysaccharides and glycoside hydrolases during seedling development of the drought-tolerant wheat cultivar (cv. Hong Mang Mai) were compared with the drought-sensitive cultivar (cv. Shirasagikomugi). After 15d of culture in water at 22 degrees C under constant irradiance of 98mumolm(-2)s(-1), the length of the coleoptile and leaf sheath of Hong Mang Mai seedlings was 1.7 times longer than those of Shirasagikomugi seedlings. In the cell walls isolated from coleoptiles and leaf sheaths of the seedling of the two cultivars, the contents of arabinose, xylose, and glucose changed during development. The cell walls were fractionated progressively with 50mM CDTA, 50mM Na(2)CO(3), 1M KOH and 4M KOH, and sugar composition was determined. The amount of CDTA-soluble fraction from the Hong Mang Mai cell walls was 2.4-fold higher than that from the Shirasagikomugi cell walls at 6d of culture, and a considerable decrease was observed during development. The ratio of arabinose to xylose in 1M KOH-soluble fraction from the two cultivars decreased. The amount of 4M KOH-soluble fraction from the Shirasagikomugi cell walls was affected much more than those of the Hong Mang Mai cell walls. Many glycoside hydrolase activities were detected in the protein fractions from coleoptiles and leaf sheaths of the two cultivars, and the activities of licheninase, 1,3-1,4-beta-glucanase, and 1,3-beta-glucanase in the LiCl-soluble protein fraction increased drastically during development of the Shirasagikomugi seedlings. These findings suggest that the metabolism of the cell wall matrix polysaccharides of the drought-tolerant wheat cultivar is far different from that of the drought-sensitive wheat cultivar during seedling development.     

Specific roles of AtEXPA1 in plant growth and stress adaptation

Expansins are a group of proteins that appear to be involved in the disruption of the noncovalent bonds within the cell wall. The distinctly expressed expansin genes can independently regulate cell expansion in place and time, and their diverse expression patterns suggest their distinct effects on plant growth. In this paper, we analyzed the effects of excessive expansin AtEXPA1 on plant growth and plant adaptation to NaCl and ABA stresses by overexpressing its gene in Arabidopsis plants. The AtEXPA1 overexpressing plants exhibited stunted shoot growth, mainly during the early phase of vegetative growth, and the growth of transgenic seedlings was also impaired. Comparing with their growth under normal growth condition, the AtEXPA1 overexpressing plants showed alleviated impairment under salt and ABA stress conditions. These results suggest that, although excessive AtEXPA1 could disturb cell wall organization and lead to growth reduction, it inversely helped enhancing cell wall organization under stress conditions and thus helped plant better to adapt to adverse environment.     

Levels of endogenous abscisic acid and indole-3-acetic acid influence shoot organogenesis in callus cultures of rice subjected to osmotic stress

Osmotic stress and endogenous hormone levels may have a role in shoot organogenesis, but a systematic study has not yet to investigate the links. We evaluated the changes of the endogenous indole-3-acetic acid (IAA) and abscisic acid (ABA) levels in rice (Oryza sativa L. cv. Tainan 5) callus during shoot organogenesis induced by exogenous plant growth regulator treatments or under osmotic stress. Non-regenerable callus showed low levels of endogenous ABA and IAA, with no fluctuation in level during the period evaluated. The addition of 100 μM ABA or 2 mM anthranilic acid (IAA precursor) into Murashige and Skoog basal induction medium containing 10 μM 2,4-D enhanced the regeneration frequency slightly, to 5 and 35%, respectively, and their total cellular ABA or IAA levels were increased significantly, correspondingly to the treatments. However, the regeneration frequency was greatly increased to 80% after treatment with 0.6 M sorbitol or 100 μM ABA and 2 mM anthranilic acid combined. Both treatments produced high levels of total cellular ABA and IAA at the callus stage, which was quickly decreased on the first day after transfer to regeneration medium. Thus, osmotic stress-induced simultaneous accumulation of endogenous ABA and IAA is involved in shoot regeneration in rice callus.     

小麦胚芽鞘扩展蛋白特性及对水分胁迫的响应

扩展蛋白是植物细胞壁延伸过程中的关键调节因子,在植物的生长发育以及对逆境的响应过程中起着重要作用。本文选用小麦(HF 9703)胚芽鞘为材料,采用Hepes法和SDS法分别提取小麦胚芽鞘扩展蛋白,通过改良的植物组织伸长测定仪测定其活性,并利用扩展蛋白抗体进行免疫印迹以检测其丰度,主要研究了小麦胚芽鞘扩展蛋白的特性及对水分胁迫的响应。结果表明:Hepes法提取的扩展蛋白活性较高,而SDS法的提取效率高;离体小麦胚芽鞘扩展蛋白的活性具有pH依赖性,且随缓冲液的交替更换(pH 4.5:pH 6.8)而反复逆转;扩展蛋白主要定位于细胞壁中;小麦胚芽鞘扩展蛋白和黄瓜下胚轴扩展蛋白具有交叉重组活性,但这种活性具有种属特异性。水分胁迫诱导小麦胚芽鞘扩展蛋白的活性和丰度提高,扩展蛋白活性的提高在小麦对水分胁迫的抗性方面可能具有重要作用。