种壳对大麦种子生理和成熟胚培养的影响及机理探讨Ⅱ

用30％PEG6000对大麦种子进行浸种等处理,并对其吸胀生理的多数指标进行了系统分析和检测,研究结果表明：种壳显著延缓种子的吸水进程;种壳对多数分子的渗漏有较大影响。N的渗漏主要来自胚乳,Mn^2 渗漏主要来自胚,种壳显著影响K^ 渗漏。去壳促使吸胀期间可溶性蛋白量和过氧化物酶同工酶的活性显著升高。吸胀使IAA、GA3含量提高,使胚乳中的ABA含量显著减少;去壳促进吸胀初期胚乳中的ABA、IAA、GA3含量,去壳减少吸胀后期胚中的ABA、IAA、GA3含量。种子去壳后吸胀24h有显著较高的萌发率、愈伤组织诱导率。     

小花草玉梅种子萌发过程中常见内源激素含量的变化及萌发机制

植物内源激素在调节种子休眠和萌发过程中具有极其重要的作用。本研究运用反式高效液相色谱(RP-HPLC)与紫外检测器联用的方法对小花草玉梅干种子、吸胀种子和露白种子中的赤霉素GAs、脱落酸ABA、玉米素ZT和生长素IAA含量进行了检测,旨在研究植物内源激素水平在种子萌发过程中的变化是否直接关系着小花草玉梅种子在光照或黑暗条件下的萌发能力。结果显示,种子吸胀过程中,光照促进了ZT同时抑制了ABA的积累,并且ZT对ABA萌发抑制作用的解除也受光的促进,露白种子中的ABA/ZT,ABA/GAs和(ABA+IAA)/(GAs+ZT)水平在黑暗条件下高于光照条件,上述均是导致光照条件下种子萌发率较高的重要原因;相对于干种子,IAA含量在种子吸胀初期急速下降,(ABA+IAA)/(GAs+ZT)在种子萌发过程中有所降低,而ABA/GAs却表现出明显的上升趋势;各激素水平所受光照的影响均在种子开始露白时显著减弱,另外,吸胀第9天是小花草玉梅种子萌发过程中激素变化的一个关键的时间节点。总之,种子萌发并非直接关系着GA含量的升高和ABA含量的降低。因光照直接促进了小花草玉梅种子的萌发,本研究认为高寒草甸充足的光照和较强的太阳辐射通过调节种子内源激素水平在一定程度上对种子萌发过程中的休眠解除具有重要作用。     

GA3和变温层积对天女木兰种子萌发及内源激素的影响

用不同质量浓度GA3浸泡天女木兰种子并结合变温层积处理,应用高效液相色谱法对不同时期种子中4种激素GA3、IAA、ABA、ZR含量进行测定,并测量种胚长和萌发率,以探讨天女木兰种胚发育,内源激素含量变化与种子休眠萌发之间的调控关系,为进一步研究种子休眠机理提供理论基础。结果表明:(1)天女木兰成熟种子胚发育不完全,胚乳内高浓度ABA和低浓度GA3是其休眠的主要原因。(2)GA3处理能促使天女木兰种子提前30d完成形态后熟,并以1 500mg·L-1 GA3处理效果最佳。(3)在变温层积过程,天女木兰种胚发育可分三个阶段:阶段Ⅰ(0~70d)完成种胚进一步分化;阶段Ⅱ(70~120d)种胚快速生长时期;阶段Ⅲ(120~150d)休眠完全解除,种子具备发芽能力。种子能否打破休眠主要取决于阶段Ⅰ和Ⅱ的状况。(4)GA3/ABA、IAA/ABA和ZR/ABA在种子后熟期间的变化同胚生长发育存在一致性,认为内源激素的相对水平对种子休眠具有重要的调控作用。     

桃儿七种子低温层积过程中胚形态及生理生化物质变化

为探究低温层积过程中桃儿七种子胚形态及生理生化变化与休眠解除的内在联系,该研究通过低温层积处理(90 d)解除桃儿七种子休眠,观测不同层积时间种子胚形态、胚率、发芽率、营养物质(淀粉、可溶性蛋白质、可溶性糖)含量、内源激素［赤霉素(GA)、吲哚乙酸(IAA)、脱落酸(ABA)］水平及呼吸途径关键限速酶［丙酮酸激酶(PK)、琥珀酸脱氢酶(SDH)、6 磷酸 葡萄糖脱氢酶(G 6 PDH)］的活性变化。结果显示：(1)在低温层积过程中,桃儿七种子胚形态为鱼雷或子叶型胚;种子发芽率在层积后期(60~75 d)显著提高(P＜0.05)。(2)层积后,种子内淀粉含量及PK活性、SDH活性显著降低(P＜0.05),其可溶性蛋白含量和IAA含量显著升高(P＜0.05),萌发促进物和抑制物比例(GA/ABA、IAA/ABA、GA+IAA/ABA)也呈升高趋势。(3)种子胚率与其可溶性糖含量呈显著负相关关系,种子发芽率与其可溶性蛋白呈显著正相关关系(P＜0.05)。研究发现,桃儿七种子无形态休眠;种子内营养物质的分解转化为种子休眠解除过程中各种代谢活动提供能量,且淀粉可能是此过程中最主要的供能物质;磷酸戊糖途径(PPP)的活化、萌发促进物和抑制物比例的升高及IAA含量的显著上升是桃儿七解除休眠的关键。     

珍稀濒危植物金丝李种子的休眠机理

珍稀濒危植物金丝李(Garcinia paucinervis)种子的萌发十分缓慢,探讨其休眠机理,可为该物种的种质资源保育与可持续利用提供理论依据。本文对金丝李种子种皮结构及其透水性,剔除部分种皮和胚乳后种子的萌发情况,胚乳和胚等粗提物的活性,储藏、层积和不同温度下种子萌发情况,萌发过程中内源激素含量等进行了研究。结果表明:金丝李种皮无栅栏细胞层,下表面的角质层较薄;种皮对种子的吸胀阻碍小;随着种孔端剔除种皮和胚乳程度的加深,金丝李种子的萌发进程逐渐延长,甚至降低其萌发率,种脐端削除处理对种子萌发影响不大;内果皮、种皮、胚乳和胚中可能存在抑制金丝李种子萌发和生长的内源抑制物;新鲜种子胚率达86.12%,低温层积后胚率无显著变化;低温层积处理延缓其萌发进程,对萌发率无显著影响,4℃低温层积是储藏金丝李种子的较好方法;种子萌发对温度敏感,在32℃培养下可打破种子休眠,萌发速度显著加快。种子萌发过程中ABA含量降低,GA与ABA、IAA与ABA的比值随种子萌发显著升高,萌发促进与抑制物比例逐渐趋于提高。因此,金丝李种子存在内源抑制物,同时缺乏萌发促进物质,导致生理休眠。该种子休眠特性使其幼苗生长能应对生境的季节变化,种群在风险环境中得以延续,避免大量幼苗竞争。植被破坏导致种子萌发阶段受阻是造成金丝李濒危的原因之一。     

南方红豆杉种子综合处理过程中内源激素的动态变化

为揭示南方红豆杉种子内源激素与休眠的关系,采用酶联免疫吸附法(ELISA)测定了经过层积处理的种皮和胚乳的脱落酸(ABA)、赤霉素(GA3 )、吲哚乙酸(IAA)、玉米素核苷(ZR)4种内源激素含量的变化情况.结果表明:种子胚乳中内源ABA的含量随着层积时间的延长而逐渐下降,GA含量增加,IAA和ZR的含量先增加后降低...     

不同种类柑橘的抗旱性及其与内源激素变化的关系

以红橘、朱橘、土橘、枳、枳橙、实生酸柚、实生甜橙、黄柑为材料,进行不同降水量的长期干旱胁迫后,测定生物量、抗旱系数及叶片中赤霉素(GA1+3)、玉米素核苷(ZR)、生长素(IAA)、脱落酸(ABA)含量,分析GA1+3、ZR、IAA、ABA的相对含量变化及ABA/GA1+3、ABA/ZR、ABA/IAA与8个种类柑橘抗旱性的关系.结果表明:在降水量低于1200 mm(对照)条件下,随降水量的减少不同种类柑橘的生物量明显降低,其抗旱系数及抗旱力大小顺序为朱橘红橘枳土橘枳橙实生酸柚实生甜橙黄柑.不同种类柑橘叶片中ABA含量随降水量的减少而增加,GA1+3、ZR和IAA含量随降水量的减少而降低.在抗旱性强的柑橘种类中,ABA/GA1+3和ABA/ZR的增加幅度大,ABA/IAA变化与种间抗旱力的关联度不大.ABA和GA1+3的相对含量与柑橘抗旱系数分别呈极显著正相关和极显著负相关,ZR的相对含量与抗旱性系数呈极显著或显著负相关,IAA的相对含量与抗旱性系数相关性不明显.对柑橘抗旱性的促进效果,以ABA最大,其次为GA1+3和ZR,IAA最小.     

枣雄性不育种质胚败育与内源激素变化的关系

以雄性不育1号和梨枣为试材,研究了枣果实发育过程内源激素变化与胚败育的关系。结果表明,种子Z、GA3和IAA含量高于果肉利于枣种子早期发育。种子发育过程Z／GA3和Z／IAA持续下降,IAA／GA3在胚败育的关键时期硬核期前后呈现双峰,且种子内的IAA／GA3大于果肉。在果实发育过程中,胚败育果实内Z、GA3和IAA含量及比值呈现出异常变化。早期胚败育果实果肉Z、GA3和IAA含量高于含种胚的果肉,呈现较高水平的Z／GA3和Z／IAA及低水平的IAA／GA3。果实发育后期瘪种子中IAA、GA3和ABA亏缺,Z异常累积,Z／GA3异常升高。     

北五味子体细胞胚发生过程中内源IAA、ABA和GA3含量的动态变化

以北五味子不同发育阶段的体细胞胚为材料,利用高效液相色谱法(HPLC)对其内源吲哚乙酸(IAA)、脱落酸(ABA)和赤霉素(GA3)含量进行了测定,探讨内源激素在体细胞胚正常发育过程中所具有的作用。在北五味子体细胞胚发生过程中,IAA和GA3含量均呈现先增加后降低趋势,ABA含量表现为先降低后增加趋势。IAA和ABA含量分别在球形胚阶段达到最大值和最小值,而GA3含量在心形胚阶段达到最大值。与正常体细胞胚相比,畸形胚中IAA和GA3含量高于各时期的正常胚状体,而ABA含量则显著低于其他各时期的正常胚状体。高水平的IAA在北五味子体细胞胚的发生阶段起重要的作用,而ABA在体细胞胚的成熟过程中起关键作用,GA3则与其体细胞胚的生长发育过程密切相关。但过高水平的IAA和GA3,以及过低水平的ABA会使体胚无法正常发育成熟,可能是引起畸形胚数量增加的主要原因之一。     

珍稀濒危植物珙桐种子休眠萌发过程中内源激素的变化

珙桐是我国特有珍稀濒危植物,休眠期长且具二次休眠现象。将处于休眠萌发过程中的珙桐种子依据胚根长度划分为4个阶段,利用高效液相色谱(HPLC)测定各阶段种子及其内果皮中ABA(脱落酸)、GA(赤霉素)、KT(细胞分裂素)、IAA(3-吲哚乙酸)4种内源激素含量,分析其比值动态变化,并与幼苗阶段进行比较。结果显示:未破壳种子的内果皮中内源激素含量以ABA最高,其次是GA、IAA、KT,随着种子破壳后四种激素含量显著降低。除ABA外,种子中GA、IAA和KT含量随着胚根的伸长逐渐升高,但仍低于幼苗阶段。此外,随着胚根伸长,种子中GA/ABA、IAA/ABA、KT/ABA比值逐渐增大,其中以GA/ABA的变化最显著。因此,珙桐种子的休眠和萌发可能主要受ABA和GA的平衡和拮抗来调控。     

水稻胚胎发生与萌发早期分离胚中内源激素的变化

以酶联免疫吸附检测技术分析了水稻(Oryza sativa ssp．iapoica)分离胚不同发育时期及萌发早期的内源激素含量的动态变化。GA1含量是所测激素中含量最高的。GA1的变化趋势基本上与ABA相反。花后4d的胚中GAl和ABA的含量最高;花后8d到18d,GA1的含量下降,而ABA含量增加。在早期萌发过程中,种子吸涨后2d的胚中GA,含量迅速上升,而ABA下降。GA1／ABA的最高比值也出现在吸涨后2d的胚中。iPAs和ZRs的最高含量也出现在开花后4d的胚中,但随后含量均下降到相当低的水平,并几乎没有变化。研究结果进一步证实了GAl在早期胚胎发生和萌发过程中起重要的作用;推测iPAs和ZRs可能仅在胚胎发生的早期起作用;GA1与ABA含量之间的相对平衡控制着胚胎发育的过程。用分离胚作为测试材料可以避免胚乳等其他组织成分的干扰,从而比较准确地反映了胚的内源激素变化。此外,本研究是首次用4d的水稻幼胚作为激素含量测定的起始材料。     

Physiological characteristics and related gene expression of after‐ripening on seed dormancy release in rice

After‐ripening is a common method used for dormancy release in rice. In this study, the rice variety Jiucaiqing (Oryza sativa L. subsp. japonica) was used to determine dormancy release following different after‐ripening times (1, 2 and 3 months). Germination speed, germination percentage and seedling emergence increased with after‐ripening; more than 95% germination and 85% seedling emergence were observed following 1 month of after‐ripening within 10 days of imbibition, compared with <45% germination and 20% seedling emergence in freshly harvested seed. Hence, 3 months of after‐ripening could be considered a suitable treatment period for rice dormancy release. Dormancy release by after‐ripening is mainly correlated with a rapid decline in ABA content and increase in IAA content during imbibition. Subsequently, GA₁/ABA, GA₇/ABA, GA₁₂/ABA, GA₂₀/ABA and IAA/ABA ratios significantly increased, while GA₃/ABA, GA₄/ABA and GAs/IAA ratio significantly decreased in imbibed seeds following 3 months of after‐ripening, thereby altering α‐amylase activity during seed germination. Peak α‐amylase activity occurred at an earlier germination stage in after‐ripened seeds than in freshly harvested seeds. Expression of ABA, GA and IAA metabolism genes and dormancy‐related genes was regulated by after‐ripening time upon imbibition. Expression of OsCYP707A5, OsGA2ox1, OsGA2ox2, OsGA2ox3, OsILR1, OsGH3‐2, qLTG3‐1 and OsVP1 increased, while expression of Sdr4 decreased in imbibed seeds following 3 months of after‐ripening. Dormancy release through after‐ripening might be involved in weakening tissues covering the embryo via qLTG3‐1 and decreased ABA signalling and sensitivity via Sdr4 and OsVP1.     

柑橘果实生长发育过程中果实不同部位的4种内源激素含量变化的研究

以单性结实的国庆1号温州蜜柑和自花结实的华农本地早橘为材料,研究了果实生长发育过程中果实不同部位的吲哚乙酸(IAA)、脱落酸(ABA)、赤霉素(GA1/3)和玉米素核苷(ZR)含量的变化。结果表明:(1)国庆1号果皮IAA、GA1/3和ZR含量在幼果阶段均相对较高,随后果皮和果肉IAA含量均趋下降,而在果实膨大期内果肉ABA和果皮、果肉GA1/3、ZR含量均出现上升峰值,果实成熟采收时果皮和果肉ABA含量均明显回升。(2)华农本地早种子、果皮和果肉IAA及其种子ABA含量均在果实膨大期内出现明显峰值,在幼果阶段至果实膨大初期内种子GA1/3和ZR含量均居较高并出现明显上升,对应的果皮、果肉4种内源激素水平均相对较低且变幅小。还就两结实类型柑橘果实生长发育与其内源IAA、ABA、GA1/3和ZR含量动态的关系进行了讨论。     

Role of the testa in preventing cellular rupture during imbibition of legume seeds

Studies with the seeds of soybean, navy bean, pea, and peanut were made to determine the extent of leakage of intracellular enzymes during imbition. Embryos with intact testae from all four species were found to leak detectable activities of either intracellular enzymes of the cytosol (glucose-6-phosphate dehydrogenase) or enzymes found in both the cytosol and organelles (malate dehydrogenase, glutamate dehydrogenase, glutamate oxaloacetate transaminase, and NADP-isocitrate dehydrogenase) after 6 hours imbition at 25 C. Pea and peanut embryos with testae leaked considerably lower levels of activity for these enzymes than did those of soybean and bean. Leakage of mitochondrial marker enzymes (fumarase, cytochrome c oxidase, and adenylate kinase) was not detected from embryos with testae, suggesting that a differential diffusion of intracellular components out of cells occurred. Soybean and bean embryos without testae leaked high, and proportionally (per cent dry seed basis) similar, levels of all cytosol, cytosol-organelle, and mitochondrial marker enzymes and protein during imbibition, indicating that cell membranes were not differential to leakage and that they had ruptured. Pea and peanut embryos without testae leaked detectable activities of all cytosol and cytosol-organelle enzymes, although fumarase was the only detectable mitochondrial marker enzyme leaked, suggesting that some degree of differential leakage may have occurred in these species. The outermost layers of embryo cells of seeds without testae of all four species absorbed and sequestered the nonpermeating pigment Evan's blue after 5 to 15 minutes imbibition, indicating that membranes had ruptured. This occurred to a much lesser extent in seeds with intact testae. Both soybean and bean embryos without testae were observed to disintegrate during imbibition, whereas those of pea and peanut did not. These data indicate that seeds of certain legumes are susceptible to cellular rupture during imbibition when seed coats are damaged or missing.     

Polyamines, IAA and ABA during germination in two recalcitrant seeds: Araucaria angustifolia (Gymnosperm) and Ocotea odorifera (Angiosperm)

Background and Aims

Plant growth regulators play an important role in seed germination. However, much of the current knowledge about their function during seed germination was obtained using orthodox seeds as model systems, and there is a paucity of information about the role of plant growth regulators during germination of recalcitrant seeds. In the present work, two endangered woody species with recalcitrant seeds, Araucaria angustifolia (Gymnosperm) and Ocotea odorifera (Angiosperm), native to the Atlantic Rain Forest, Brazil, were used to study the mobilization of polyamines (PAs), indole-acetic acid (IAA) and abscisic acid (ABA) during seed germination.

Methods

Data were sampled from embryos of O. odorifera and embryos and megagametophytes of A. angustifolia throughout the germination process. Biochemical analyses were carried out in HPLC.

Key Results

During seed germination, an increase in the (Spd + Spm) : Put ratio was recorded in embryos in both species. An increase in IAA and PA levels was also observed during seed germination in both embryos, while ABA levels showed a decrease in O. odorifera and an increase in A. angustifolia embryos throughout the period studied.

Conclusions

The (Spd + Spm) : Put ratio could be used as a marker for germination completion. The increase in IAA levels, prior to germination, could be associated with variations in PA content. The ABA mobilization observed in the embryos could represent a greater resistance to this hormone in recalcitrant seeds, in comparison to orthodox seeds, opening a new perspective for studies on the effects of this regulator in recalcitrant seeds. The gymnosperm seed, though without a connective tissue between megagametophyte and embryo, seems to be able to maintain communication between the tissues, based on the likely transport of plant growth regulators.     

A method for profiling classes of plant hormones and their metabolites using liquid chromatography-electrospray ionization tandem mass spectrometry: an analysis of hormone regulation of thermodormancy of lettuce (Lactuca sativa L.) seeds

A highly selective and sensitive method for the simultaneous analysis of several plant hormones and their metabolites is described. The method combines high-performance liquid chromatography (HPLC) with positive and negative electrospray ionization-tandem mass spectrometry (ESI-MS/MS) to quantify a broad range of chemically and structurally diverse compounds. The addition of deuterium-labeled analogs for these compounds prior to sample extraction permits accurate quantification by multiple reaction monitoring (MRM). Endogenous levels of abscisic acid (ABA), abscisic acid glucose ester (ABA-GE), 7'-hydroxy-abscisic acid (7'-OH-ABA), phaseic acid (PA), dihydrophaseic acid (DPA), indole-3-acetic acid (IAA), indole-3-aspartate (IAAsp), zeatin (Z), zeatin riboside (ZR), isopentenyladenine (2iP), isopentenyladenosine (IPA), and gibberellins (GA)1, GA3, GA4, and GA7 were determined simultaneously in a single run. Detection limits ranged from 0.682 fmol for Z to 1.53 pmol for ABA. The method was applied to the analysis of plant hormones and hormonal metabolites associated with seed dormancy and germination in lettuce (Lactuca sativa L. cv. Grand Rapids), using extracts from only 50 to 100 mg DW of seed. Thermodormancy was induced by incubating seeds at 33 degrees C instead of 23 degrees C. Germinating seeds transiently accumulated high levels of ABA-GE. In contrast, thermodormant seeds transiently accumulated high levels of DPA after 7 days at 33 degrees C. GA1 and GA3 were detected during germination, and levels of GA1 increased during early post-germinative growth. After several days of incubation, thermodormant seeds exhibited a striking transient accumulation of IAA, which did not occur in seeds germinating at 23 degrees C. We conclude that hormone metabolism in thermodormant seeds is surprisingly active and is significantly different from that of germinating seeds.     

Phytohormones and structure of cells ofAcer saccharinum seed embryo

Endogenous phytohormone levels and cell structure ofAcer saccharinum embryos were studied during seed development. Mature seeds had high water content (50%) and were able to germinate immediately after fruit abscission. The submicroscopic cell structure was similar to the structure of functionally active cells. Free and conjugated indole-3-acetic acid (IAA), abscisic acid (ABA), zeatin riboside and dihydrozeatin content and gibberellin-like substances (GLS) activity were determined during embryo maturation. Decrease in ABA, free IAA and cytokinin levels was observed at the end of maturation. Mature seeds contained considerable amounts of conjugated IAA and had high GLS activity.