Isolation and Characterization of Ty1/copia-like Reverse Transcriptase Sequences from Mung Bean (Vigna radiata)

Ty1/copia-like sequences were amplified from mung bean (Vigna radiata (L.) Wilczek) genomic DNA, by PCR with degenerate oligonucleotide primers corresponding to highly conserved domains in the Ty1/copia-like retrotransposons. PCR fragments of roughly 270 bp were isolated and cloned, and forty clones were sequenced. Thirty-six of the forty clones had unique nucleotide sequences, and eighteen clones had a frameshift, a stop codon, or both. Alignment of the nucleotide sequences indicated that these clones, denoted Tvr, fell into nine subgroups and nine ungrouped sequences. The nucleotide sequence similarity between these elements ranged from 8% to 100%, which indicates high level of sequence heterogeneity among these clones. A phylogenetic analysis comparing these clones with corresponding sequences from other plant species showed that some of the Tvr clones are more closely related to Ty1/copia-like retrotransposons from other species than to other Tvr clones. Dot blot analysis revealed that Ty1/copia-like retrotransposons comprise about 9.3% of the mung bean genome.     

表油菜素内酯对绿豆上胚轴内源IAA及其氧化酶的影响

用0.5ppm表油菜素内酯处理绿豆幼苗,显著促进上胚轴伸长生长,若切除真叶则可抑制表油菜素内酯诱导的效应。三碘苯甲酸(TIBA)也可抑制表油菜素内酯促进的伸长生长。外源IAA能部分恢复TIBA的抑制效应。经处理的上胚轴内源IAA含量明显高于对照。暗示表油菜素内酯可能通过对内源IAA的调节来促进绿豆上胚轴的伸长生长。 表油菜素内酯处理的绿豆上胚轴组织中,与生长素降解密切相关的IAA氧化酶以及过氧化物酶活性均明显低于对照。     

芦丁对绿豆幼苗营养生长的影响及其与IAA的相互作用

观察了植物体内的天然黄酮芦丁和吲哚乙酸(IAA)对绿豆幼苗营养生长的影响并测定胚轴中的芦丁和IAA含量.光照条件下芦丁(60μg/mL以下)处理对绿豆幼苗生长有一定促进作用,表现为胚轴和主根伸长加快、侧根数目增多、鲜重或干重增加;而光照条件下更高浓度芦丁(80μg/mL以上)处理及黑暗条件下芦丁(20～100μg/mL)处理对绿豆幼苗生长有抑制作用.当培养基中的芦丁浓度为60～80 μg/mL时,光照下的幼苗比暗处理的幼苗在胚轴中积累更多的芦丁;而芦丁浓度为40μg/mL以下和接近100μg/mL时幼苗在光照下累积的芦丁较暗处理的幼苗更少.0.1μg/mL以上的IAA促进芦丁的累积而进一步抑制幼苗胚轴和主根的伸长.当培养基中含有40 μg/mL的芦丁和0.5μg/mL的IAA时,胚轴中累积的芦丁达到高峰.芦丁降低黄化幼苗内源性IAA在胚轴中的累积,并抑制幼苗对IAA的吸收.     

表油菜素内酯对绿豆上胚轴核酸代谢的影响

0.5～5 ppm放线菌素D和0.01～0.1ppm亚胺环己酮均能抑制epiBR促进的绿豆上胚轴的生长。DNA、RNA含量测定表明epi—BR处理促进了它们的含量增加。绿豆上胚轴经 epiBR处理48h后,可增强其RNA聚合酶活性(以~H—UTP掺入RNA的DPM计算);DNA和RNA水解酶的活性随着epiBR处理时间的延长而逐渐降低;TIBA则抑制epiBR的上述作用。     

切除上胚轴对绿豆子叶衰老逆转及其核酸和蛋白质代谢的效应

在绿豆子叶衰老达到不归点(萌发后5～6d)前切除上胚轴可使开始衰老的子叶中核酸和蛋白质含量回升,衰老短期逆转。衰老不归点后的子叶中核酸和蛋白质变化的主要特征是:丧失了较多的核主带DNA、25S、18S rRNA以及大部分可溶性蛋白质组分,一种小分子DNA 组分完全消失。不归点前切除上胚轴可使上述核酸和蛋白质组分明显增加,表明子叶衰老的逆转可能与这些重要功能物质的回升有关。在切除上胚轴的茎顶涂抹IAA,能阻止子叶核酸和蛋白质回升,也消除了切除上胚轴对子叶裹老的逆转作用。     

绿豆bHLH转录因子家族的鉴定与生物信息学分析

bHLH是真核生物中重要的一类转录因子,其主要由碱性氨基酸区和螺旋-环-螺旋区组成。本研究利用生物信息学的方法鉴定到122个绿豆bHLH转录因子,并对其理化性质、保守结构域、基因结构、在染色体上的分布、系统进化以及部分典型基因的组织表达差异等进行分析。结果表明,bHLH转录因子理化性质差异较大;含有2个保守结构域,分别位于N端的碱性氨基酸区和C端的螺旋-环-螺旋区,碱性氨基酸区含有His5-Glu9-Arg13保守序列,与靶基因结合有关,HLH区含有Arg23和Arg55,与形成二聚体有关,同时含有5种保守元件;bHLH基因在11条染色体上分布不均匀,5号、7号和8号染色体上分布较多,1号、4号和10号染色体上分布较少,大部分基因含有1~9个不等的内含子,在染色体上成簇状分布;122个bHLH转录因子可分为11个亚家族。多数bHLH基因在绿豆根、茎、叶、花和种子等组织中均有表达,但具有组织表达特异性,且不同基因表达量差异较大。本研究为进一步研究绿豆bHLH转录因子家族的生物学功能奠定基础。     

Bradyrhizobium elkanii, Bradyrhizobium yuanmingense and Bradyrhizobium japonicum are the main rhizobia associated with Vigna unguiculata and Vigna radiata in the subtropical region of China

Cowpea (Vigna unguiculata) and mung bean (Vigna radiata) are important legume crops yet their rhizobia have not been well characterized. In the present study, 62 rhizobial strains isolated from the root nodules of these plants grown in the subtropical region of China were analyzed via a polyphasic approach. The results showed that 90% of the analyzed strains belonged to or were related to Bradyrhizobium japonicum, Bradyrhizobium liaoningense, Bradyrhizobium yuanmingense and Bradyrhizobium elkanii, while the remaining represented Rhizobium leguminosarum, Rhizobium etli and Sinorhizobium fredii. Diverse nifH and nodC genes were found in these strains and their symbiotic genes were mainly coevolved with the housekeeping genes, indicating that the symbiotic genes were mainly maintained by vertical transfer in the studied rhizobial populations.     

蓝光对绿豆下胚轴愈伤组织形成和生长过程中蛋白质代谢的影响

蓝光、白光和黑暗对绿豆下胚轴愈伤组织形成和生长过程中蛋白质代谢的影响不同。培养后３～１８ ｄ ,蓝光处理材料的可溶性蛋白质含量明显高于白光处理,更高于黑暗培养的材料。蓝光和白光明显促进３Ｈ亮氨酸掺入蛋白质,而蓝光和白光处理后游离氨基酸含量与黑暗对照相比,下降时间早,幅度大。在培养过程中,蛋白酶活性的变化与游离氨基酸相似。蛋白质合成抑制剂环己酰亚胺（ＣＨＭ） 抑制愈伤组织生长,其中以蓝光最大,白光次之,黑暗最小。在培养基中加入ＣＨＭ 愈早,抑制程度愈大。实验表明,ＣＨＭ 抑制愈伤组织蛋白质合成,也是以蓝光最甚。由此可见,蓝光促进绿豆下胚轴愈伤组织的形成、生长和蛋白质合成。     

Enhancing effects of heat shock and gibberellic acid on the thermotolerance in etiolated Vigna radiata. I. Physiological aspects on thermotolerance

The effect of heat shock on the thermotolerance of etiolated mung bean seedlings ( Vigna radiata L. cv. Wilczek) and the effects of gibberellic acid (GA) were studied. The potentially lethal temperature of etiolated mung bean seedlings was 45°C. But, when seedlings were pretreated with a heat-shock period at 40°C for 1 h before incubation at 45°C, they become thermotolerant and survived the 45°C treatment. The addition of actinomycin D or cycloheximide during the heat-shock period decreased the subsequent thermotolerance of the seedlings. Depending upon the time of its application, GA appeared to have multiple effects: (1) when applied during the 40°C heat-shock period, GA enhanced the heat-shock effect; (2) when applied during the 45°C potentially lethal temperature period, GA enhanced the subsequent growth of hypocotyls. This suggests that GA makes the seedlings tolerant to the potentially lethal temperature; (3) when GA was applied during a following 25°C growth period to seedlings which had been exposed first to 40°C and then 45°C, it promoted growth, suggesting that GA enhanced the restoration of the seedlings from high temperature damage. The role of GA and heat shock in the acquisition of thermotolerance in etiolated mung bean seedlings are discussed.     

Responses of Vigna radiata to Foliar Application of 28-Homobrassinolide and Kinetin

Effects of 28-homobrassinolide (HBR) and kinetin (KIN) on photosynthesis, nitrogen metabolism, and the seed yield were studied. The leaves of 25-d-old plants of Vigna radiata (L.) Wilczek were sprayed with 0.01, 1.0 or 100 M aqueous solution of KIN, or 0.0001, 0.01 or 1.0 M that of HBR. KIN and especially HBR increased the activities of nitrate reductase and carbonic anhydrase, chlorophyll and total protein contents and net photosynthetic rate in the leaves, and pod number and seed yield, at harvest.     

NO和NADPH-黄递酶在绿豆下胚轴不定根发生和发育过程中的变化

研究了一氧化氮(NO)供体普钠(SNP)、一氧化氮清除剂C-PTIO和一氧化氮合酶(NOS)抑制L-NAME对绿豆(Vigna radiataL.)下胚轴插条生根的影响.并对不定根生期间手条基部NO 和NADPH-黄递酶的时空变化进行了检测.所试浓度SNP均明显促进下胚轴不根发生.分别插条切取后24h和36h于其基部维管束之间检测到NADPH-黄递酶(NOS标记酶)阳性反应和NO荧光,根原基也于48h在相同位置出现,并于60h进一步伸长.48~60h期间,NADPH、黄递的阳性反应及NO荧光有增强趋势,并主要分布在不定根分生组织中.L-NAME既减弱NADPH-黄递酶的阳性反应和NO荧光,也延缓不不定根发生;而c-PTIO对NO荧光及不定根生均有抑制作用.上述结果证明:NO在不定根发生及发育过程中有重要作用,而且此过程中的NO很可能由类似的NOS催化产生.     

Effects of Uniconazole on Leaves Photosynthesis,Root Distribution and Yield of Mung Bean (Vigna radiata)

Journal of Plant Growth Regulation - Uniconazole was a plant growth retardant with effect of regulating plant growth and development, however, there were very few studies on its application to mung...     

蚓粪中的植物激素及其对绿豆插条不定根发生的促进作用

利用高效液相色谱证实了环毛蚓（Pheretima sp.)蚓粪中至少存在两种植物激素（IAA和GA3）,蚓粪中IAA和GA3的含量分别可达13.9mg/kg和49.9mg/kg。为原土的13.6和8.2倍,采用包括蚓粪提取液,原土提取液,Haogland营养液,蒸馏水,IAA,GA3等在内的11种供试液进行绿豆插条不定根发生的生物试验,结果表明,在相同条件下,蚓粪提取液处理的绿豆插条比其它处理提早3天出现不定根,统计证明,与原土提取液,Haogland营养液,蒸馏水等处理相比,蚓粪提取液处理可显提高绿豆插条不定根的数量,长度和生物量,蚓粪提取液对不定根发生数量的诱导效应与85-170μmol/L的IAA溶液相当,但从不定根根长和根生长量来看,蚓粪提取液的效果优于IAA。     

NO和NADPH-黄递酶在绿豆下胚轴不定根发生和发育过程中的变化(英文)

研究了一氧化氮(NO)供体硝普钠(SNP)、一氧化氮清除剂c-PTIO和一氧化氮合酶(NOS)抑制剂L-NAME对绿豆(Vigna radiata L.)下胚轴插条生根的影响,并对不定根发生期间插条基部NO和NADPH-黄递酶的时空变化进行了检测。所试浓度SNP均明显促进下胚轴不定根发生。分别在插条切取后24 h和36 h于其基部维管束之间检测到NADPH-黄递酶(NOS标记酶)阳性反应和NO荧光,根原基也于48 h在相同位置出现,并于60 h进一步伸长。48-60h期间,NADPH-黄递酶的阳性反应及NO荧光有增强趋势,并主要分布在不定根分生组织中。L-NAME既减弱NADPH-黄递酶的阳性反应和NO荧光,也延缓不定根发生;而c-PTIO对NO荧光及不定根发生均有抑制作用。上述结果证明:NO在不定根发生及发育过程中有重要作用,而且此过程中的NO很可能由类似的NOS催化产生。     

Individual and interactive effects of temperature,carbon dioxide and abscisic acid on mung bean (Vigna radiata) plants

We studied the effects of temperature, carbon dioxide and abscisic acid on mung bean (Vigna radiata). Plants were grown under 26/22°C or 32/28°C (16?h?light/8?h?dark) at 400 or 700?μmol?mol^?1 CO₂ and received ABA application of 0 or 100?μl (10?μg) every other day for three weeks, after eight days of initial growth, in growth chambers. We measured 24 parameters. As individual factors, in 16 cases temperature; in 8 cases CO₂; in 9 cases ABA; and as interactive factors, in 4 cases, each of temperature?×?CO₂, and CO₂?×?ABA; and in 2 cases, temperature?×?ABA were significant. Higher temperatures increased growth, aboveground biomass, growth indices, photochemical quenching (qP) and nitrogen balance index (NBI). Elevated CO₂ increased growth and aboveground biomass. ABA decreased growth, belowground biomass, qP and flavonoids; increased shoot/root mass ratio, chlorophyll and NBI; and had little role in regulating temperature–CO₂ effects.

Abbreviations: A_N: net CO₂ assimilation; E: transpiration; F_v/F_m: maximum quantum yield of PSII; g_s: stomatal conductance; LAR: leaf area ratio; LMA: leaf mass per area; LMR: leaf mass ratio;φPSII: effective quantum yield of PSII; qNP: non-photochemical quenching; qP: photochemical quenching; SRMR: shoot to root mass ratio; WUE: water use efficiency     

外源赤霉素、6-苄基腺嘌呤及矿物质对水培绿豆芽生长的影响

以‘中绿一号’绿豆为原料用水培法制作豆芽,研究赤霉素、6-苄基腺嘌呤和矿物质对其生长的影响,优化绿豆芽营养液的配方。结果表明,使用由100mg·L-1赤霉素、5mg·L-16-苄基腺嘌呤、0.810g·L-1硝酸钾、0.950g·L-1硝酸钙、0.500g·L-1硫酸镁复配而成的营养液可以缩短豆芽生长周期,且培养出的绿豆芽芽长适宜（4.25cm±0.44cm）、根长较短（1.15cm±0.23cm）、轴径较粗（2.78cm±0.16mm）,芽体洁白而脆嫩。     

Interaction of paclobutrazol and indole-3-butyric acid in relation to rooting of mung bean (Vigna radiata) cuttings

Paclobutrazol (PB) only slightly stimulated the rooting of mung bean cuttings but, interestingly, the number of adventitious roots formed was dramatically increased when PB was used together with indole-3-butyric acid (IBA). Application of PB in the first phase of root formation, when root initials are induced, caused the greatest enhancement of the promotive effect of IBA on rooting. Investigation of the effect of PB on uptake, transport and metabolism of [5^-3H]-IBA in mung bean cuttings revealed some changes in the rate of metabolism of IBA in comparison with control cuttings. PB was found to be involved in the partitioning of carbohydrates along the cuttings. Application of sucrose, like PB to the base of IBA-treated cuttings enhanced the effect of IBA. The patterns of the effects of PB and IBA, separately and together, on rooting were similar in defoliated and intact cuttings, however the number of roots was much lower in the defoliated cuttings, which lacked a source of assimilates. PB counteracted the effect of GA₃ in the upper regions of the cuttings and seemed to increase the sink capacity at the base of the cuttings. The results of the present study clearly demonstrated the enhancing influence of PB on IBA stimulation of the rooting of mung bean cuttings. It is suggested that PB may affect the rate of metabolism of IBA during rooting and the status of the local sink, in the base of the cuttings, thus partially contributing to the enhancement of the rooting-promotive effect of IBA.     

NaCl胁迫下SA浸种绿豆幼苗的生长及生理特征

以'中绿一号'绿豆品种为材料,对不同浓度水杨酸(SA)浸种绿豆种子在NaCl胁迫条件下的萌发、幼苗生长及相关生理指标变化进行分析.结果显示:(1)与未胁迫对照相比,未浸种对照绿豆在100～500 mmol·L-1 NaCl处理下的发芽率、芽长、根长和叶片叶绿素含量显著降低,而幼苗叶片丙二醛(MDA)与脯氨酸(Pro)含量水平显著上升(P＜0.05),且其升降幅度随NaCl胁迫浓度提高而增加;(2)与未浸种对照相比,未胁迫对照种子的萌发和幼苗的生长在20 mg·L-1 SA处理中受到抑制,而在40～80 mg·L-1 SA处理条件下得到促进,至100 mg·L-1 SA时又受到显著抑制.(3)适当浓度的SA浸种能够显著提高盐胁迫下绿豆幼苗的芽长、根长、叶绿素的含量,降低了MDA和Pro含量;在NaCl胁迫浓度为100～300 mmol·L-1时的SA适宜浓度浸种为60 mg·L-1,而500 mmol·L-1 NaCl时为80 mg·L-1.研究表明,适当浓度的SA浸种能有效缓解盐胁迫对绿豆幼苗的伤害,提高其耐盐性.     

Effect of NaCI on nitrate reductase,glutamate dehydrogenase and glutamate synthase inVigna radiata calli

The effect of NaCI stress on the activities of nitrate reductase (NR), glutamate dehydrogenase (GDH) and glutamate synthase (GOGAT) in callus lines ofVigna radiata which differ in salt resistance, was studied at weekly intervals upto 28 d of growth. After 28 d, the NaCI resistant callus (selected at 300 mM NaCI) at NaCI concentrations higher than 200 mM maintained higher NR activity than non-selected line. NaCI stress also affects aminating and deaminating activities of GDH. The NADH-GDH activity in the presence of NaCI was higher in the resistant than non-selected line. On the other hand, NAD-GDH activity in both the lines was completely inhibited after 7 d of growth. The increased activity of NADH-GDH in resistant calli may play a vital role in protecting the cells from toxic effect of increased endogenous level of ammonia which probably accumulates due to efficient NO₃ ⁻ reduction. NADH-GOGAT activity was found to decrease under salt stress in both the callus lines. Nitrogen assimilation in salt-resistant calli under salt stress was found to be characterized by high NR and NADH-GDH activities, concomitantly with low GOGAT activity. The authors are grateful to DST and CSIR for financial assistance.     

H2O2在黄瓜和绿豆下胚轴不定根形成中的作用

以黄瓜和绿豆下胚轴插条为试验材料,研究了H2O2对不定根形成的诱导作用,以及在此过程中与IAA、NO的关系。结果表明,H2O2能促进不定根的形成,最佳浓度是100μmol/L;H2O2的清除剂———过氧化氢酶(CAT)抑制了IAA诱导的不定根形成,说明H2O2可能介导了IAA诱导不定根形成的过程;NO清除剂———PTIO能抑制H2O2诱导的不定根形成,表明NO在H2O2诱导不定根形成的过程中起介导作用。综合上述结果,推测在不定根形成的过程中可能存在以下关系:IAA首先诱导H2O2合成,H2O2升高后,再诱导NO产生,最终导致不定根生成。