Physiological and biochemical changes associated with cotton fibre development. II. Auxin oxidising system

Changes in IAA oxidase, and in cytoplasmic and ionically wall-bound peroxidase activities were studied in the developing fibres of three cotton cultivars ( Gossypium hirsutum L. cv. Gujarat-67, cv. Khandwa-2 and G. herbaceum L. cv. Digvijay), designated as long, medium and short staple cultivars, respectively. In all the three cultivars IAA oxidase activity was low during the fibre elongation phase, while the activity increased significantly during the secondary thickening phase. The increase in IAA oxidase activity in the three cultivars showed close correspondence with their respective total period of elongation. No relationship between cytoplasmic peroxidase activity and fibre development was discernible. The ionically bound wall peroxidase activity, however, recorded low levels during the elongation phase and higher levels during the secondary thickening phase. The role of wall peroxidase in cessation of elongation growth is discussed.     

Changes in o-Diphenol Oxidase During Fibre Development in Cotton

Quantitative and electrophoretic changes in o-diphenol oxidase(o-diphenol: O₂ oxidoreductase, E.C. 1 10,3.1) were studiedduring the entire period of cotton (Gossypium arboreum L. cv.Sanjay) fibre development. A significant increase in o-diphenoloxidase activity was recorded during the fibre initiation phaseand it is suggested that a shift in redox balance towards oxidationmay play an important role in fibre initiation. Low o-diphenoloxidase activity during elongation and its high activity duringthe phase of secondary thickening, together with isoenzyme patterns,suggest an important role of this enzyme in cotton fibre development.The role of o-diphenol oxidase in relation to auxin turnoverand redox balance is discussed. Gossypium arboreum, cotton, fibre development, o-diphenol oxidase, redox balance, auxin turnover     

棉纤维细胞伸长生长与过氧化物酶和IAA氧化酶的关系

棉纤维细胞于开花当天从棉胚珠表皮上发生,随即开始伸长生长,星S型生长曲线。棉纤维细胞的可溶性蛋白、过氧化物酶活性和IAA氧化酶活性同伸长生长的关系不大;而离子型结合的细胞壁蛋白质含量、过氧化物酶活性和IAA氧化酶活性同棉纤维细胞的伸长生长关系较大,表现在棉纤维细胞快速伸长期活性较低,而在伸长生长停止时出现活性高峰,同棉纤维细胞的伸长生长有负相关现象。     

Effects of plant growth regulators on secondary wall thickening of cotton fibres

The effect of plant growth regulators on the secondary wall thickeningof cotton fibre was studied. The results indicated that the GA_S andiP+iPA levels in the fibre of field-grown cotton plantsremained almost constant but the IAA and ABA levels changed considerably duringfibre development. Although the change in both IAA and ABA levels seemed not tobe closely related with the rate of cellulose accumulation, there was still arelationship between the ratio of ABA to IAA and secondary wall thickening. Inin vitro studies, ABA (50mol·L^–1) markedly enhanced theaccumulation of dry matter and cellulose in the fibre cell wall duringsecondarywall thickening, but no similar effect was observed with NAA, GA₃ orkinetin treatments. The role of ABA in secondary wall thickening of cottonfibreis discussed.     

棉花纤维发育过程中内源激素含量和纤维品质变化及其关系研究

以新疆棉区优质棉品种‘新陆早16号’、品质中等品种‘新陆早10号’和‘新陆早13号’以及品质较差品种‘02-DB’为材料,测定了棉纤维发育过程中内源生长素(IAA)、赤霉素(GA4)、玉米素(ZR)和脱落酸(ABA)含量和主要纤维品质指标的变化,分析内源激素含量变化与纤维品质形成的关系。结果表明:不同品种棉花纤维发育中纤维内源激素变化趋势基本相似,其差异主要表现在IAA、GA4、ZR和ABA的含量大小及峰值出现的时间方面。‘新陆早16号’在纤维发育前期有较高IAA、GA4、ZR含量和较低的ABA含量,表现出纤维伸长速率较高、快速伸长时期较长等特征;而且在次生壁加厚期ZR峰值出现较早,有利于棉纤维成熟,从而表现出较优的纤维品质。‘02-DB’在纤维发育前期由于ABA含量较高影响了纤维伸长速率和快速伸长期的时间,同时后期ZR峰值出现晚,使纤维发育受到影响,而最终品质较差。可见,在棉花纤维伸长期IAA、GA4、ZR含量高而ABA含量低、次生壁加厚期ZR峰值出现早则有利于优质棉纤维形成。     

Role of peroxidase and esterase activities during cotton fiber development

Cytoplasmic and salt-extracted wall peroxidase and nonspecific esterase activities along with growth analysis were investigated during the entire period of cotton fiber development. Both the peroxidase fractions, when assayed with chlorogenic and ferulic acids as substrates, recorded low levels during the fiber elongation phase, and a close relationship between cessation of elongation growth and increase in peroxidase activity was discernible. Nonspecific esterase activity in both cytoplasmic and salt-extracted fractions, on the other hand, showed higher activity during the elongation phase, whereas during the secondary thickening phase it decreased. The role of cytoplasmic peroxidase in IAA oxidation is discussed. It is suggested that esterases and peroxidases associated with wall fractions may well be involved in turnover of phenolic acids that are cross-linked to wall polysaccharides.     

Physiological and biochemical changes associated with cotton fibre development

Growth kinetics and levels of auxin substances were studied in three cotton cultivars, designated as long, medium and short staple cultivars. Fibre length and dry weight plotted against boll age showed sigmoidal patterns and were fitted to a logistic curve by computer curvilinear regression analysis. The final length of the fibre in different cultivars was the product of the rate of elongation per day and the total period of elongation. Further, considerable overlap between the elongation and the secondary thickening phases was recorded.
No relationship between auxin substances and rate of fibre elongation was discernible. The peak levels of auxin substances in all the cultivars were recorded before or about the time when elongation had just started, and it is concluded that the auxin synthesized during the elongation phase is consumed in elongation growth. Thus there is necessarily no relationship between remaining auxin and growth.     

Esterase ontogeny in cotton fibre

Growth parameters and esterase isoenzyme patterns were studied in developing cotton fibre. Fibre length and dry weight when plotted against boll age showed four distinct stages; (i) initiation, (ii) elongation, (iii) secondary thickening, and (iv) maturation. Stage specificity and changing intensities of esterase isoenzyme suggested a finely modulated control of gene expression during fibre development.     

Effects of Guaianolide derivative I on fibre initiation and elongation in Gossypium arboreum L. in relation to adverse effects of cool temperature

Cotton ovules collected during late September with prevalent night cool temperature (15°C), cultured at 30°C/15°C i.e. cycling temperatures in Beasley and Ting medium had very few epidermal cells showing bulging. Supplementing cultures with guaianolide derivative I (E-13 methyldehydrocostus lactone) promoted fibre initiation. At—1 day preanthesis (DPA), IAA oxidase activity declined in guaianolide-treated cultures but increased during the elongation phase and was enhanced during the secondary wall thickening phase. However, o-diphenol oxidase activity was adversely affected during the fibre initiation phase. The activities of all the other enzymes studied viz. acid invertase, phenylalanine ammonia lyase, -glucosidase and IAA oxidase increased, except -galactosidase, during the later phase in comparison with the controls. The present study indicates that guaianolide derivative I triggers early initiation and promotes fibre elongation by regulatin o-diphenols and IAA-oxidation levels, which in turn check wall loosening. Considerable enhancement in the soluble acid invertase activity by this compound suggests its role in apoplastic sucrose hydrolysis, thereby preventing its accumulation.Abbreviations DPA days pre anthesis - DAC days after culturing - DAA days after anthesis     

Physiological and Biochemical Changes Associated with Cotton Fibre Development. IV. Glycosidases and {beta}-1,3-Glucanase Activities

Developing cotton fibre was analysed from 12 days post anthesis(DPA) till maturity for the activity of wall degrading enzymes,ß-galactosidase, ß-glucosidase, -mannosidaseand ß-1,3-glucanase. Each enzyme was estimated inthree different fractions namely cytoplasmic, ionically wall-boundand covalently wall-bound. There was a significant correlationbetween ß-galactosidase and ß-glucosidaseactivities in the covalently bound fraction, and the rate offibre elongation. Similarly, covalently bound ß-1,3-glucanaseactivity showed an increasing trend up to 18 DPA, i.e. aboutthe time when maximum rate of fibre elongation was achieved. The results presented here suggest that covalently wall-boundglycosidases may have an importafit role in cell wall loosening.Earlier reports providing evidence against the involvement ofthese enzymes in elongation growth in intact system, may perhapsbe due to scant attention paid to the subcellular distributionof these enzymes. Gossypium hirsutum, cotton fibre, glucanase, glycosidase, wall-loosening     

A peptide hormone gene,GhPSK promotes fibre elongation and contributes to longer and finer cotton fibre

Cotton fibres, the single‐celled trichomes derived from the ovule epidermis, provide the most important natural material for the global textile industry. A number of studies have demonstrated that regulating endogenous hormone levels through transgenic approaches can improve cotton fibre qualities. Phytosulfokine‐α (PSK‐α) is a novel peptide hormone in plants that is involved in regulating cell proliferation and elongation. However, its potential applications in crop genetic improvement have not been evaluated. In this study, we describe how exogenous PSK‐α application promotes cotton fibre cell elongation in vitro. Chlorate, an effective inhibitor of peptide sulfation, suppressed fibre elongation in ovule culture. Exogenously applied PSK‐α partly restored the chlorate‐induced suppression. A putative PSK gene (GhPSK) was cloned from Gossypium hirsutum. Expression pattern analysis revealed that GhPSK is preferentially expressed in rapidly elongating fibre cells (5–20 days postanthesis). Overexpression of GhPSK in cotton increased the endogenous PSK‐α level and promoted cotton fibre cell elongation, resulting in longer and finer fibres. Further results from electrophysiological and physiological analyses suggest that GhPSK affects fibre development through regulation of K⁺ efflux. Digital gene expression (DGE) profile analysis of GhPSK overexpression lines indicates that PSK signalling may regulate the respiratory electron‐transport chain and reactive oxygen species to affect cotton fibre development. These results imply that peptide hormones are involved in cotton fibre growth and suggest a new strategy for the biotechnological improvement of cotton fibre quality.     

Potential Role of Abscisic Acid in Cotton Fiber and Ovule Development

Fibers and ovules of a cotton cultivar (Gossypium hirsutum L. Trambak-108) were analyzed for growth and free abscisic acid (ABA) content by indirect enzyme immunoassay. An inverse correlation between fiber elongation and ABA content was observed. In the seed, accumulation of ABA was observed during secondary thickening and the maturation phase. The potential role of ABA in fiber and seed development is discussed. Received June 25, 1997; accepted October 15, 1997     

Involvement of wall microtubules in gibberllin promotion and kinetin inhibition of stem elongation

The elongation of light-grown azuki bean (Azukia angularis =Vigna angularis) epicotyl segments was promoted by indoleaceticacid (IAA) and this IAA-induced elongation was inhibited byboth kinetin and benzimidazole (BIA). Increased stem thickeningwas observed with kinetin- or BIA-treated segments, but thiswas not accompanied by incresed cell number in the transversedirection, suggesting that both kinetin and BIA promoted lateralcell expansion. Colchicine at a concentration with no effecton IAA-induced elongation reversed both the kinetic- and BIA-inducedinhibition. Electron-microscopic examination revealed that wall microtubulesin cells treated with kinetin together with IAA ran parallelto the cell axis, while wall microtubules in cells treated withonly IAA were randomly oriented. In the cell treated with gibberellintogether with IAA, wall microtubules ran tranverse to the cellaxis. (Received July 13, 1973; )     

In Vitro Phosphorylation of Proteins in IAA-Treated Mesocotyls in Zea mays

Five-mm sections of elongation zones of Zea mesocotyls wereincubated for designated periods with various concentrationsof IAA. In vitro protein phosphorylation in the soluble fraction(85,000 x g supernatant) prepared from the sections was analyzedby sodium dodecyl sulfate-polyacrylamide gel electrophoresis.The phosphorylation of proteins in the soluble fraction thathad been prepared from sections incubated for 20 min in thepresence of 10{small tilde}^s M IAA was greater than that inthe sections incubated for 20 min without IAA. The amount ofphosphorylation of proteins per protein became higher when higherconcentrations increased (10{small tilde}⁸—10{small tilde}⁵M).The growth of sections incubated in the presence of 10{smalltilde}⁸ M IAA or higher concentrations was greater than thatof sections incubated in the absence of IAA. The promotion ofgrowth by IAA was greater at higher concentrations of IAA. Proteinsin the soluble fraction, prepared from sections incubated for20 min in the presence of 10{small tilde}⁵ M IAA, were phosphorylatedin the presence of either 10 fM cAMP, 10 µM cGMP, 100µM W-7, 100 µM W-5, 20 µM H-7 or 20 µMHA1004. The calmodulin antagonist, W-7, and the inhibitor ofprotein kinase C, H-7, inhibited the phosphorylation of proteinsstimulated by incubation with IAA. These results suggest thatIAA promotes cell elongation via protein phosphorylation thatdepends on calmodulin-dependent protein kinase and protein kinaseC. (Received November 29, 1995; Accepted May 20, 1996)     

Physiological and biochemical changes associated with cotton fiber development. VIII. Wall components

Fibers of three cotton cultivars (Gossypium hirsutum L.) H-4, H-8 and (G. arboreum) G. Cot-15, which shows variation in staple length were analyzed for growth in terms of fiber length and fresh and dry mass. From the growth analysis cotton fiber development is divided in four distinct phases i.e. (i) initiation (ii) elongation (iii) secondary thickening and (iv) maturation. Rate of fiber elongation and rate of water content shows close parallelism. Highly esterified and less esterified pectic fraction along with high and low molecular weight xyloglucan fractions were estimated from fiber walls of all the three cotton genotypes. Xyloglucans were fractioned in to high and low molecular weight by alkali treatment, 1 M and 4 M KOH respectively. Xyloglucan content shows inverse correlation with fiber elongation. Role of water content and wall components in determination of staple length in cotton genotypes is discussed.     

The Role of Auxin in the Apical Regulation of Leaf Abscission in Cotton (Gossypium hirsutum L.)

The petiole abscission induced by deblading cotyledonary leavesof cotton (Gossypium hirsutum L. cv. Delta Pine) was acceleratedby the presence of the intact shoot apex or, in decapitatedplants and explants, by application to the stem (proximal application)of indol-3yl-acetic acid (IAA) or 1-aminocyclopropane-l-carboxylicacid (ACC). IAA and ACC accelerated the abscission of debladedpetioles whether applied above or below the cotyledonary node.Transport of IAA to the node was not required for the responseto proximal IAA. [2,3-¹⁴C]ACC was readily transported to thenodal region whether applied to the stem above or below thenode. Application of IAA or ACC to the stem did not induce theabscission of intact leaves or of debladed petioles treateddistally with IAA The acceleration of abscission by proximal IAA, but not thatcaused by ACC, was prevented if explants were treated with a-aminooxyaceticacid (AOA), an inhibitor of ACC-synthase. AOA also preventedthe acceleration of abscission caused by the shoot apex. Theprogress of abscission in debladed explants was greatly delayedby silver thiosulphate (STS—an inhibitor of ethylene action),whether or not the explants were treated with IAA or ACC. Itis suggested that the speeding effects of the shoot apex andof proximal auxin on the abscission of debladed petioles requiresauxin-induced ACC synthesis. The possibility is discussed thatACC may function as a mobile abscission promoter Key words: Abscission, ACC, ACC-synthase, cotton (Gossypium), proximal auxin     

Interaction of a Brassinosteroid with IAA and GA3 in the Elongation of Cucumber Hypocotyl Sections

A synthetic brassinosteroid, 22,23(S,S)-homobrassinolide (hBR),was examined for its interaction with IAA and GA₃ in the elongationof hypocotyl sections of light-grown cucumber (Cucumis salivusL. cv. Aonagajibai) seedlings. hBR alone was less active thanIAA. Its optimal concentration was around 10 µM and thelowest effective concentration between 10 and 100 µM,which is more than 100 times higher than that of brassinolide.hBR was more active in sections from younger seedlings. Itsgrowth-promoting effect was negated or greatly reduced by inhibitorsof auxin-induced elongation such as p-chlorophenoxyisobutyricacid and kinetin. hBR acted synergistically with IAA and 2,4-Dbut not with GA₃ showing only an additive effect. Sequentialtreatment of sections with hBR and then with IAA also resultedin synergistic enhancement of auxininduced elongation, but whenthe order of treatment was reversed, hBR was inactive. The synergisticeffect was obtained with 1 h pretreatment with hBR and couldbe reduced by subsequent washing with water. There was no sequentialinteraction between hBR and GA₃. The synergistic pretreatmenteffects of hBR and GA₃ were simply additive to each other. Amembrane-bound ATPase inhibitor, dicyclohexylcarbodiimide, inhibitedthe hBR-induced elongation, but did not affect GA₃-induced elongation.The findings led to the conclusion that brassinosteroids enhanceauxin action and possess growth-promoting activity which isindependent of that of gibberellin. (Received November 9, 1984; Accepted February 18, 1985)