黄瓜贴接苗砧木子叶淀粉代谢的动态变化特征

该研究以黄瓜品种‘中农18号’、南瓜砧木品种‘京欣砧5号’为试验材料,以南瓜自根苗(P)和去除1片子叶及生长点的南瓜苗( /P)为对照,采用单子叶贴接法进行黄瓜/南瓜异体嫁接(C/P)和南瓜/南瓜自体嫁接(P/P),测定嫁接后砧木子叶形态指标和淀粉代谢的动态变化,分析嫁接后去除砧木子叶对嫁接苗生长发育的影响,以揭示砧木子叶淀粉代谢在黄瓜嫁接苗生长中的作用,为黄瓜嫁接苗的壮苗培育提供理论依据。结果显示：(1)贴接后,C/P、P/P和 /P砧木子叶鲜质量和面积显著增大,且增加量依次递减,表现为 /P > C/P > P/P > P。(2)贴接后,C/P和P/P砧木子叶中淀粉含量在嫁接后0~3 d时降低,之后迅速升高,至嫁接后13 d再次逐渐降低,且C/P砧木子叶淀粉含量及其淀粉分支酶(SBE)和水解酶(β AL)活性均显著高于P/P。(3)在嫁接后0~10 d 去除砧木子叶可显著抑制C/P嫁接苗接穗和根系生长,减弱根系活力,同时降低根系可溶性糖含量及其CWIN、HXK基因表达水平,并以嫁接后0 d 去除砧木子叶的抑制效果最显著。研究表明,黄瓜C/P单子叶贴接苗中,砧木子叶作为贮存器官,在幼苗生长早期以淀粉形式储存光合产物,之后淀粉水解成单糖为嫁接苗接穗和根系快速生长提供物质和能量。     

The Role of Cytokinins and Ethylene in Bean Cotyledon Senescence. The Effect of Free Radicals

During ageing of bean (Phaseolus vulgaris L.) cotyledons in plants with modified life span the time-course of four cytokinins, ethylene, and the end products of free radical attack, lipofuscin-like pigments (LFP), were studied. UV irradiation shortened cotyledon life span, while epicotyl decapitation prolonged it. In controls, LFP increased at the senescence onset but at the end of life span it returned to the initial level. Ethylene increased more than 3-fold at the time of abscission. The content of individual cytokinins (zeatin, zeatin riboside, isopentenyl adenine, isopentenyl adenine riboside) varied differently during ageing but they did not decreased in any case under level observed in young cotyledons at the time of abscission. UV irradiation resulted in 14-fold increase in LFP concentration at the end. Ethylene increased 8-fold 2 h after irradiation. Individual cytokinins increased after UV irradiation to a different extent and time-course, nevertheless cotyledon life span was shortened. Decapitation induced LFP decrease. On day 13, LFP abruptly increased and than decreased and stayed lowered until abscission. Ethylene was maximum on day 24, at the time of abscission, it was above 200 % of control. Decapitation produced transient decrease in some cytokinins namely zeatin and isopentenyl adenine riboside.     

Sequencing and Expression of the Gene that Encodes a 20-kDa Polypeptide of the PSI Complex from Cucumber Cotyledon

A cDNA clone encoding the polypeptide from cucumber PS I thatmigrates with an apparent molecular weight of 20 kDa on SDS-polyacrylamidegels has been isolated. The 907-bp sequence of this clone hasbeen determined and contains one large open reading frame thatencodes a 22,720-Da precursor polypeptide (207 amino acid residues).The molecular weight of the mature polypeptide was predictedto be 17,037-Da (153 amino acid residues). The deduced aminoacid sequence of this protein indicates that it is routed towardsthe stromal side of the thylakoid membrane and has no membrane-spanningregions. The sequence also confirmed the identity of the proteinas the product of the psa D gene. Chemical cross-linking offerredoxin to the PS I complex identified the 20-kDa subunitas the ferredoxin-binding protein. Northern hybridization experimentsrevealed that the mRNA of approximately 1,100 nucleotides forthe 20-kDa polypeptide was present in etiolated cucumber cotyledons,and its level increased about 5-fold during greening. The 20-kDapolypeptide was not detected by immunoblotting in etiolatedcotyledons, and it accumulated only after illumination. Labelingexperiments in vivo showed the absence of incorporation of [³⁵S]Metinto the polypeptide in etiolated cotyledons. These resultssuggest that the expression of the psa D gene is controlledat the translational level. (Received April 5, 1990; Accepted June 28, 1990)     

The Staurotypus Turtles and Aves Share the Same Origin of Sex Chromosomes but Evolved Different Types of Heterogametic Sex Determination

Reptiles have a wide diversity of sex-determining mechanisms and types of sex chromosomes. Turtles exhibit temperature-dependent sex determination and genotypic sex determination, with male heterogametic (XX/XY) and female heterogametic (ZZ/ZW) sex chromosomes. Identification of sex chromosomes in many turtle species and their comparative genomic analysis are of great significance to understand the evolutionary processes of sex determination and sex chromosome differentiation in Testudines. The Mexican giant musk turtle (Staurotypus triporcatus, Kinosternidae, Testudines) and the giant musk turtle (Staurotypus salvinii) have heteromorphic XY sex chromosomes with a low degree of morphological differentiation; however, their origin and linkage group are still unknown. Cross-species chromosome painting with chromosome-specific DNA from Chinese soft-shelled turtle (Pelodiscus sinensis) revealed that the X and Y chromosomes of S. triporcatus have homology with P. sinensis chromosome 6, which corresponds to the chicken Z chromosome. We cloned cDNA fragments of S. triporcatus homologs of 16 chicken Z-linked genes and mapped them to S. triporcatus and S. salvinii chromosomes using fluorescence in situ hybridization. Sixteen genes were localized to the X and Y long arms in the same order in both species. The orders were also almost the same as those of the ostrich (Struthio camelus) Z chromosome, which retains the primitive state of the avian ancestral Z chromosome. These results strongly suggest that the X and Y chromosomes of Staurotypus turtles are at a very early stage of sex chromosome differentiation, and that these chromosomes and the avian ZW chromosomes share the same origin. Nonetheless, the turtles and birds acquired different systems of heterogametic sex determination during their evolution.     

Culture of Cotyledon and Hypocotyl Protoplasts from True Potato Seedlings in Solanum tuberosum L.

Culture of protoplast using cotyledon and hypocotyl as the donor tissue from true potato seedlings (TPSs) of 3 breeding lines (DTO-33, ND 860-2 and BN 9815-3) of Solanum tuberosum L. was studied. The cotyledons and hypocotyls of TPSs just extended were excised and digested in an enzyme solution containing 1 % cellulase and 0. 5 % macerozyme for 17—20 h after vacuum infiltration of the tissue in the solution. The protoplasts were cultured in an improved liquid medium and transferred onto solid media for callus culture and shoot regeneration. Some factors affecting the efficiency of cotyledon and hypocotyl protoplast culture were studied. The results showed that using the cotyledons and hypocotyls as donor tissues for protoplast isolation and culture in potato, the division frequency of protoplast derived cells was significantly higher than that using the leaves and shoot-tips of the test-tube plantlets: the yield and quality of the protoplast from TPSs cultured under continuous high light intensity (3000 Ix) were much higher than the TPSs cultured under low light intensity (1000 Ix), and no intact protoplast was ever obtained from the TPSs cultured in continuous dark condition. Vacuum infiltration of the cotyledon and hypocotyl segments in enzyme solution before digestion increased protoplast yield. The yield of protoplasts from hypocotyl tissue was significantly higher than from the cotyledon, but there was no significant difference in quality between the protoplast derived from the two tissues. The significance, advantages and shortcomings of using the cotyledons and hypocotyls as the donor tissues for isolation and culture of potato protoplasts are dicussed.     

Separation of the Effects of Photoperiod and Hormones on Sex Expression in Cucumber

The effects of photoperiod on the sex expression of cucumberflowers were examined in relation to the endogenous GA and ethyleneproduction with short day (SD)- and long day (LD)-sensitivecultivars under controlled environmental conditions. Growth of plants was greatly reduced under LD in both typesof cultivars. Reflecting this, GA activity was greater and ethyleneevolution less in plants grown under SD than in those underLD. However, the number of pistillate flowers increased underSD and LD in SD- and LD-sensitive cultivar, respectively, indicatingthat a change in sex expression did not coincide with hormonalchanges in the SD-sensitive cultivar. Thus, the effect of photoperiodon the sex expression was found to be separate, at least inpart, from the effect of GA or ethylene in cucumber. (Received September 16, 1982; Accepted December 7, 1982)     

Ca2+对黄瓜子叶离体培养物花芽分化的影响

黄瓜子叶离体培养物分化培养 0～ 8d期间 ,添加Ca2 有利于花芽分化 ,花芽分化率可从Ca2 0mmol/L的 (7.9± 5 .6 ) %上升到Ca2 6mmol/L的(31.7± 4.0 ) % ;0～ 2d和 2～ 4d无钙脉冲处理不利于花芽分化 ,高钙脉冲处理的花芽分化率比对照略高 ;4～ 6d和 6～ 8d高钙脉冲不利花芽分化 ,而无钙脉冲处理使花芽分化率上升很多。尤其是在 4～ 6d ,高钙处理使花芽分化率从 (2 2± 1.5 ) %下降到 (15 .7±3 .5 ) %。而无钙处理使花芽分化率从 (2 2 .4± 1.4) %上升到 (4 3± 3 .5 ) %。表明 0～ 8d期间不同时间段对Ca2 的需求是有差别的。相关性分析表明 :0～ 8d期间外源Ca2 影响花芽分化率与总芽中花芽比例极显著相关 ,提示Ca2 可能影响子叶向花芽或营养芽分化的趋势。本文结合已报道的黄瓜子叶培养物花原基形成的时程 ,分析了Ca2 对花原基形成和分化的影响     

Involvement of the Cotyledon in Adventitious Root Initiation in Impatiens balsamina L. Seedlings

Adventitious root primordia are found in the pre-hypocotyl tissueof developing seeds of Impatiens balsamina L. by the third weekafter petal drop, and are present in the mature seed. Aftergermination, the adventitious roots emerge from a collet swellingon the hypocotyl of the young seedlings. Removal of the colletduring the first five days results in the formation of anotherat the base of the remaining hypocotyl. Older seedlings respondto the excision of the collet by producing one or more rootsnear the cut end, unless the cut is made close to the cotyledon,when, even in nine-day seedlings, a reduced collet is formedassociated with four or fewer roots. The influence of the cotyledonon collet/root regeneration diminishes in older seedlings andin these is manifested only in hypocotyl tissue adjacent tothat organ. Impatiens balsamina, balsam, cotyledon, adventitious roots, collet     

Role of Ethylene in Avocado Fruit Development and Ripening: I. FRUIT DROP

A survey of avocado (Persea americana Mill.) fruit drop andethylene production was carried out during fruit development.Natural drop of avocado fruitlets started soon after set (May)and continued at a gradually decreasing rate until September,except for a temporarily increasing rate in late July. Fruitletsweighing up to 0?2 g dropped at a rate of over 30 percent perweek. With larger fruits, the rate was under 1 percent per week.Fruit drop ceased after September, when fruit growth declinedand the seed coat began to shrivel. A positive correlation was found between the rate of fruitletand fruit drop and ethylene production. Fruitlets with defectiveseeds produced ethylene at a very high rate of 7–10 timesmore than apparently normal fruits. The high incidence of defectiveseeds might be the cause of the very high levels of ethyleneproduction by young avocado fruitlets. The seed was found to be the main site of ethylene productionin fruitlets. Abscisic acid (ABA) levels in young abscissing fruits were 7times as high as those in non-abscissing fruits.     

Ca2+对黄瓜子叶离体培养物花芽分化的影响

黄瓜子叶离体培养物分化培养0～8d期间,添加Ca2+有利于花芽分化,花芽分化率可从Ca2+0mmol/L的(7.9±5.6)%上升到Ca2+6mmol/L的(31.7±4.0)%;0～2d和2～4d无钙脉冲处理不利于花芽分化,高钙脉冲处理的花芽分化率比对照略高;4～6d和6～8d高钙脉冲不利花芽分化,而无钙脉冲处理使花芽分化率上升很多。尤其是在4～6d,高钙处理使花芽分化率从(22±1.5)%下降到(15.7±3.5)%。而无钙处理使花芽分化率从(22.4±1.4)%上升到(43±3.5)%。表明0～8d期间不同时间段对Ca2+的需求是有差别的。相关性分析表明0～8d期间外源Ca2+影响花芽分化率与总芽中花芽比例极显著相关,提示Ca2+可能影响子叶向花芽或营养芽分化的趋势。本文结合已报道的黄瓜子叶培养物花原基形成的时程,分析了Ca2+对花原基形成和分化的影响。     

外源亚精胺对盐胁迫下黄瓜幼苗生长和可溶性蛋白表达的影响

采用营养液水培方法,研究了叶面喷施亚精胺(Spd)对黄瓜幼苗生长和可溶性蛋白的影响.结果表明,外源Spd可使盐胁迫黄瓜幼苗株高、茎粗、叶面积、根长和根表面积显著增加,可溶性蛋白含量明显提高,部分可溶性蛋白的表达量减弱;而对正常生长条件下黄瓜幼苗各生长指标无明显影响,但显著提高了根系中可溶性蛋白的含量.随着Spd处理时间的延续,盐胁迫幼苗至少有7种分子量分别约为67、61.5、50、47、43、29和16 kD的可溶性蛋白表达量发生明显变化,尤其使61.5、47和43 kD蛋白表达量明显减弱,50 kD蛋白甚至消失,这4种蛋白可能与外源Spd缓解黄瓜幼苗盐胁迫伤害效应关系密切.     

Effect of Abscisic Acid on Banana Fruit Ripening in Relation to the Role of Ethylene

Abstract The role of abscisic acid (ABA) in banana fruit ripening was examined with the ethylene binding inhibitor, 1-methylcyclopropene (1-MCP). ABA (0, 10⁻⁵, 10⁻⁴, or 10⁻³ mol/L) was applied by vacuum infiltration into fruit. 1-MCP (1 μL/L) was applied by injecting a measured volume of stock gas into sealed glass jars containing fruit. Fruit ripening, as judged by ethylene evolution and respiration associated with color change and softening, was accelerated by 10⁻⁴ or 10⁻³ mol/L ABA. ABA at 10⁻⁵ mol/L had no effect. The acceleration of ripening by ABA was greater at 10⁻³ mol/L than at 10⁻⁴ mol/L. ABA-induced acceleration of banana fruit ripening was not observed in 1-MCP treated fruit, especially when ABA was applied after exposure to 1-MCP. Thus, ABA's promotion of ripening in intact banana fruit is at least partially mediated by ethylene. Exposure of ABA-treated fruit to 0.1 μL/L ethylene for 24 h resulted in increased ethylene production and respiration, and associated skin color change and fruit softening. Control fruit (no ABA) was unresponsive to similar ethylene treatments. The data suggest that ABA facilitates initiation and progress in the sequence of ethylene-mediated ripening events, possibly by enhancing the sensitivity to ethylene. Received 29 January 1999; accepted 16 January 2000     

Effects of Volatiles Evolved from Excised Plant Tissues and Lemon Oil on the Growth of Mung Bean Seedlings

Volatiles evolved from excised segments of leaves of oleander(Nerium oleander) and cedar (Cedrus deodara), peels of lemon(Citrus limon), orange (Citrus sinensis) and lime (Citrus aurantifolia),and roots of wasabi (Wasabia japonica) inhibited growth of mungbean seedlings. The volatiles evolved from lemon oil also markedlyinhibited elongation of mung bean seedlings. The active constituent(s)in the volatiles were distinct from ethylene. The mode of inhibitionby the volatiles from lemon oil was different from that of ethylene.The volatiles from lemon oil alleviated the inhibitory effectof ethylene on the growth of seedlings. The volatiles from lemonoil supressed the formation of lamellar structure in chloroplasts,thus resulting in etiolation of seedlings. (Received October 25, 1982; Accepted December 29, 1982)     

Acetylcholinesterase from oat Seedlings. I. Preliminary biochemical characterization of the enzyme

The activity of acetylcholinesterase (AChE) isolated from coleoptiles of etiolated oat seedlings is strongly inhibited by neostigmine and less so by eserine. The optimum of the enzyme activity occurs at pH 7.2 and a temperature of + 36 °C. The enzyme Michaelis constant is 280 μM. Choline within the range of concentration from 0.001 to 10 mM does not affect the enzyme activity. Calcium ions at 5 mM concentration cause inhibition, while magnesium and manganese ions do not affect the enzyme activity. AChE isolated from oat seedlings differs in a number of properties from AChE occurring in the tissues of other plants. This research was supported in part by grant CPBP 05.02.4.07.     

Relationship between Ethylene and Spermidine in the Leaves of Glycyrrhiza uralensis Seedlings under Root Osmotic Stress1

The correlation between ethylene (ETH) production and spermidine (Spd) content was studied in the leaves of Glycyrrhiza uralensis seedlings under root osmotic stress. After 4-h root osmotic stress, Spd inhibited ETH production significantly while ETH had no significant influence on Spd content in leaves. After 24-h root osmotic stress, ETH production and Spd content showed significant negative correlations in leaves, and the negative correlations were still significant even if the common precursor S-adenosylmethionine (SAM) was abundant. This result suggested that the significant negative correlations were not mainly caused by the competition for the common precursor SAM. The results also showed that, after 24-h root osmotic stress, ETH enhanced while Spd reduced the production rate of H₂O₂ and markedly in leaves; furthermore, exogenous H₂O₂ accelerated the increase in ETH production and the decrease in Spd content caused by deep stress. So it could be concluded that reactive oxygen species played important roles in the significant negative correlations in the deeply stressed leaves of G. uralensis seedlings.     

Action Mechanism of Ethylene in the Control of Sugar Translocation in Relation to Rice Coleoptile Growth I. Sucrose Metabolism

The fate of ¹⁴C-glucose fed through scutella of rice (Oryzasativa L. cv. Sasanishiki) seedling explants was investigatedin relation to ethylene action on sugar translocation to growingcoleoptiles and leaves. In the scutellum, sucrose, UDPglucoseand F6P were rapidly labeled, and sucrose-phosphate synthaseactivity was higher than sucrose synthase activity. Radioactivesucrose soon appeared in both coleoptiles and leaves, and increasedrapidly. Its specific activity in both tissues became almostequal to that in the scutella. The specific activities of ¹⁴C-glucosein both coleoptiles and leaves changed almost in parallel tothose of ¹⁴C-fructose. These results suggest that sucrose wassynthesized in the scutellum and exported to the coleoptileand leaf, where it was cleaved to glucose and fructose. Ethylene slightly increased the specific activities of ¹⁴C-sucrosein all tissues, but markedly increased those of ^l4C-glucoseand -fructose only in the coleoptile. We assume that the ethyleneenhancement of sucrose transport from scutellum to the coleoptileresults from the activation of sucrose unloading in the growingcoleoptile where imported sucrose is cleaved into glucose andfructose. (Received May 25, 1987; Accepted October 30, 1987)