Adventitious shoot regeneration from Begonia × erythrophylla petiole sections is developmentally sensitive to light quality

The influence of light quality on organogenesis in vitro was investigated using Begonia  ×  erythrophylla petiole explants. Pre-treatment of in vitro donor plants by growth in the dark or under far-red or blue light reduced their competence for shoot formation when compared with those grown under red or white light. Culture of competent petiole explants under far-red, blue light or in the dark reduced the number of shoots produced per explant compared to those cultured under red or white light. Explants were found to be developmentally sensitive to both far-red and blue light, because meristem, but not primordia development was inhibited. In addition, blue light inhibition of shoot formation is not mediated directly through phytochrome, as few shoots formed on explants cultured under a mixture of red and blue light which resulted in a high P _fr/ P _tot (0.82) and would allow shoot formation in the absence of blue light. Unlike the inhibitory influence of far-red light, which is reversible, exposure to blue light permanently reduces an explant's competence for shoot formation. Our results suggest that phytochrome and an independent blue light photoreceptor, possibly a cryptochrome, can regulate shoot production from B. erythrophylla petiole explants.     

Changes of free, soluble conjugated and bound polyamine titers of jojoba explants under sodium chloride salinity in vitro

Jojoba (Simmondsia chinensis L.) single node explants were cultured in a basal medium supplemented with 17.8 microM 6-benzyladenine and four levels of sodium chloride concentration (0, 56.41, 112.82 and 169.23 mM). The free, the soluble conjugated and the insoluble bound forms of polyamines (PAs) (putrescine (Put), spermidine (Spd) and spermine (Spm)) were determined monthly during a 3-month proliferation stage. Free Put and Spd were found in higher levels in the control treatment, while Spm content was higher in the salt treatments. All soluble conjugated PAs were found to be in lower concentrations in explants growing on medium supplemented with salt, while the opposite was true for the insoluble bound PAs. It appeared that certain PAs and PAs forms could play a significant role in the adaptation mechanism of jojoba under saline conditions.     

Light quality influences adventitious shoot production from cotyledon explants of lettuce (<Emphasis Type="Italic">Lactuca sativa</Emphasis> L.)

Summary The influence of light quality on competence and determination for organogenesis was investigated using lettuce cotyledon explants. Lettuce seedlings from four genotypes were germinated in the dark or under white, red, or blue light. Cotyledon explants were excised and cultured on a shoot-inducing medium for 28 d under white light. Germination in the dark reduced shoot numbers, suggesting that light improves the competence of explants for organogenesis. When explants from seedlings germinated under white light were cultured under different light qualities, blue was found to inhibit shoot production while red light either promoted production or had no effect on shoot number compared to controls. Treatment with blue plus red light failed to overcome the inhibition by blue light. To ascertain the temporal responses of explants to light quality, they were cultured under red or blue light prior to transfer to the alternate treatment. Exposure to blue light within 7 d of excision permanently reduced explant competence for organogenesis. Exposure after this time had a minimal effect. These results suggest that both phytochrome and cryptochrome can regulate shoot production from lettuce cotyledon explants and blue light can only inhibit organogenesis, in lettuce, during a relatively small developmental window.     

NaCl胁迫对菜用大豆种子多胺代谢的影响

采用蛭石栽培,在100 mmol·L-1NaCl胁迫下,对耐盐性不同的两个品种菜用大豆种子的丙二醛(MDA)含量和多胺(PAs)代谢进行了研究.结果表明:NaCl胁迫显著增加了菜用大豆种子的MDA含量,但耐盐品种‘绿领特早’(LL)的增幅低于盐敏感品种‘理想高产95-1’(LX).与LX相比,LL种子在整个NaCl胁迫期间均维持了相对较高的游离态精胺(Spm)、结合态Spm、结合态亚精胺(Spd)、束缚态Spd和束缚态腐胺(Put)含量,较高的(Spd +Spm )/Put 和(cPAs+bPAs)/fPAs值及较低的Put/PAs值,在胁迫中、后期(9～15 d)维持了相对较高的游离态Spd含量;胁迫期间,LL的精胺酸脱羧酶(ADC)长时期(6～15 d)保持相对较高的活性,而多胺氧化酶(PAO)则长时期(6～15 d)维持相对较低的活性.综上,LL具有较强的多胺合成能力及较强的Put向Spd和Spm以及游离态多胺向结合态和束缚态多胺转化的能力,进而有效抑制了细胞的膜脂过氧化,这可能是其耐盐性较强的重要原因之一.     

Endogenous polyamine content during in vivo maturation and in vitro culture of maize pollen

Changes in polyamine content during in vivo maturation and in vitro culture of maize (Zea mays L.) pollen were studied. The endogenous content of free, conjugated and bound polyamines was analyzed during 30 days of pollen evolution, in both developmental pathways (microsporogenesis and androgenesis). The induction of androgenesis from cold-pretreated uninucleate pollen results, in most of cases, in a lower total polyamine content than that of the in vivo uninucleate pollen. These differences indicate that polyamine metabolism is altered during the induction of androgenesis, and this could be a consequence of increased polyamine assimilation. In general, pollen stages that involve cell division (tetrades, pre-anthesis pollen and four-day cultured pollen) are characterized by a predominance of free Spd. The increase of Spd and Spm in 15-day cultured pollen, when the first embryoids are formed, outline the possible implication of these polyamines in embryogenetic processes. Furthermore, these findings may contribute to the improvement of maize androgenesis yield, especially in recalcitrant genotypes, by the exogenous application of polyamines or polyamine-inhibitors to the culture medium.Abbreviations PAs polyamines - Put putrescine - Spd spermidine - Spm spermine - S free polyamine fraction - SH conjugated polyamine fraction - PH bound polyamine fraction     

枣品种耐盐性与叶片生物膜结合态多胺水平和H~＋-ATP酶活性的关系

120mmol·L^-1NaCl胁迫30d,耐盐性强的‘金丝小枣’叶片细胞质膜、液泡膜共价结合态腐胺（Put）、亚精胺（Spd）、精胺（Spm）含量及多胺（PAs）总水平与对照无显著性差异,但耐盐性弱的‘冬枣’叶片质膜共价结合态Put、Spd、Spm含量和PAs总水平及液泡膜Spd含量均显著降低;‘金丝小枣’叶片类囊体膜共价结合态Put含量、PAs总水平较对照显著降低,‘冬枣’则是Put、Spd、Spm含量及PAs总水平均显著降低。盐胁迫下,‘金丝小枣’叶片细胞质膜、液泡膜、类囊体膜非共价结合态Put、Spd、Spm含量及PAs总水平下降,但其中仅类囊体膜Spd含量显著低于对照,而‘冬枣’的3种膜上非共价结合态的这些多胺及其总水平均显著低于对照。与对照相比,盐胁迫下耐盐性不同的2个枣品种,叶片细胞质膜、液泡膜和类囊体膜H＋-ATP酶活性均降低,但降低幅度因枣品种和生物膜种类不同而异,且H＋-ATP酶活性与相应膜结合态多胺水平存在极紧密的正相关关系。结果表明,膜结合态多胺参与枣品种耐盐性的表达,调节盐胁迫下枣叶细胞中溶质的跨膜运输。     

Effect of end-of-day irradiations on polyamine accumulation in petal cultures of Araujia sericifera

We have studied photoperiodic control and the effect of phytochrome photoconversion at the end-of-day (EOD) on polyamine (PA) accumulation in petal explants of Araujia sericifera . Petals from immature flowers were cultured under long (LD) and short (SD) days. Light was provided by Gro-lux fluorescent lamps (90–100 µmol m⁻² s⁻¹). Red (R), far red (FR), red followed by far-red (R-FR) and far-red followed by red (FR-R) light treatments were applied daily at the end of the photoperiod. The free and bound putrescine (Put), spermidine (Spd) and spermine (Spm) fractions in petal explants were determined 40 days after the beginning of the culture. We also aimed to clarify the involvement of PA changes by using two inhibitors of PA biosynthesis: D- l -α-difluoromethylarginine (DFMA) and methylglyoxal bis (guanylhydrazone) (MGBG). We found PA accumulation to be under photoperiodic control, and the inhibitory effect of DFMA on this accumulation suggests that arginine decarboxylase (ADC) is the major pathway for Put biosynthesis. Polyamine levels were higher under LD, mainly as a result of the accumulation of free and bound Put. FR-EOD treatment, which dramatically reduced the R : FR ratio after LD, increased the accumulation of PA, mainly as free Put and free and bound Spd. Sequential R-FR and FR-R-EOD treatments strongly increased bound Spd. The concentration of MGBG used increased total PA accumulation, mainly as Put. However, all EOD light treatments dramatically reduced Put accumulation in the presence of MGBG. This may be due to a dual role of FR light in PA accumulation: (1) FR per se stimulates PA production, probably via ADC, and (2) in the presence of MGBG, FR inhibits Put accumulation, probably via ethylene production.     

盐胁迫对大麦幼苗质膜,液泡膜上共价和非共价结合多胺含量的影响

用不同浓度ＮａＣｌ处理７ｄ龄大麦（Ｈｏｒｄｅｕｍ ｖｕｌｇａｒｅＬ．）幼苗３ｄ。以非共价键和共价键形式分别与质膜和液泡膜微囊及膜蛋白结合的多胺含量受低 度盐的促进而被高浓度盐所抑制。以非共价键形式与膜微囊结合的各种多胺中亚精胺（Ｓｐｄ）含量最高,占膜上多胺总量的４０％－７０％,与膜蛋白共价结合的各种多胺中腐胺（Ｐｕｔ）含量占主导地位,占膜蛋白上多胺总量的３５％－６０％。在根系液泡膜上发现一种含量丰     

Variations in endogenous polyamine content of maize calli obtained from zygotic and androgenetic embryos

A comparative study of polyamine (putrescine, spermidine and spermine) levels was conducted with maize calli originating from a) immature embryos and b) pollen embryos capable of plant regeneration. The differences observed in the studied parameters of the two kinds of calluses are related to their cellular origin and to their regeneration capacity. Moreover, only the calluses proceeding from immature embryos differentiated into preembryogenic structures, which eventually developed into plants. Although total polyamine levels in pollenderived calluses were significantly higher than those from immature embryos, spermidine and spermine were the predominant polyamines in both culture types. Furthermore, polyamine fractions of these calluses also showed differences. All these phenomena may be related with the differences observed in the callus embryogenic response. These findings may be useful in understanding the implication of polyaminesin embryogenetic processes.Abbreviations IEC immature-embryo calluses - PAs polyamines - PEC pollen-embryo calluses - PH insoluble conjugated PA fraction - Put putrescine - S free PA fraction - SH soluble conjugated PA fraction - Spd spermidine - Spm spermine 2,4d-2,4 dichlorophenoxyacetic acid     

Effects of Salinity Stress on the Levels of Covalently and Noncovalently Conjugated Polyamines in Plasma Membrane and Tonoplast Isolated from Barley Seedlings

When the 7-d old barley ( Hordeum vulgare L. ) seedlings were treated with different concentrations of NaC1 for 3 d, the levels of the noncovalemly conjugated polyamines (PAs) in the plasma membrane and tonoplast vesicles and the covalently conjugated PAs in the membrane proteins were promoted by NaC1 of low concentrations and suppressed by NaC1 of high concentrations. Among the noncovalenfly conjugated PAs in the vesicles, spennidine (Spd) level was the most abundant, while putrescine (Put) content was predominant among the covalenfly conjugated PAs, accounted for 40%– 70%, 35% – 60%, respectively. In addition, the TLC (thin-layer chromatography) profiles of the benzoylated PAs presented an unknown polyamine with Rf = 0.92 (X0.92), which conjugated covalently and noncovalently in root tonoplast and its content changed as well as Spd with NaC1 treatment. The total PA contents in the roots were higher than that in the leaves, and the types and contents of covalenfly and noncovalently conjugated PAs in the tonoplast were higher than those in the plasma membrane. The results showed that the above two PAs associated with the membrane might be essential in salt adaption of cells and the maintenance of membrane function.     

多胺浸种改善盐胁迫大麦根系液泡膜功能的机理

研究了 0 .1mmol/L腐胺 (Put)和 0 .5mmol/L亚精胺 (Spd)浸种对 2 0 0mmol/LNaCl胁迫下大麦 (HordeumvulgareL .)幼苗生长速率、干物质积累、离子分布、液泡膜蛋白结合多胺含量以及液泡膜膜脂组分与功能的影响。结果表明 ,Put和Spd浸种均可缓解盐胁迫对大麦幼苗的盐害 ,促进生长和干物质积累 ,降低大麦幼苗体内 [Na ]/[K ]。与盐处理的对照植株相比 ,Put和Spd浸种均可提高大麦幼苗根系液泡膜磷脂含量 ,降低糖脂结合半乳糖含量 ,而膜上非共价结合多胺含量Spd PAx (一种未知多胺 )与Put Dap (二氨基丙烷 )之比 ( (Spd PAx) / (Put Dap) )、共价和非共价结合多胺总量均上升。统计分析结果表明 ,液泡膜非共价结合多胺 (Spd PAx) / (Put Dap)与H _ATPase和H _PPase活性呈显著正相关关系。     

Effect of exogenous spermidine on polyamine metabolism in water hyacinth leaves under mercury stress

The influence of exogenous spermidine (Spd) on arginine decarboxylase (ADC), ornithine decarboxylase (ODC), polyamine oxidase (PAO) activities and polyamines (PAs), proline contents in water hyacinth leaves under Mercury (Hg) stress was investigated after 6 days treatment. The results showed that free putrescine (Put) content increased, the contents of free spermidine (Spd) and spermine (Spm) and the (Spd + Spm)/Put ratio in water hyacinth leaves decreased significantly with the increase of the Hg concentrations. Hg stress also disturbed the activities of ADC, ODC and PAO and caused changes on proline content. Compared to the Hg-treatment only, exogenous Spd (0.1 mM) significantly reduced the accumulation of free Put, increased the contents of free Spd and Spm and the ratio of (Spd + Spm)/Put in water hyacinth leaves. Furthermore, exogenous Spd enhanced the activities of ADC, ODC and PAO and significantly increased proline content. The PS-conjugated PAs and PIS-bound PAs changed in the same trend as free PAs. These results suggest that exogenous Spd can alleviate the metabolic disturbance of polyamines caused by Hg in water hyacinth leaves.     

多胺浸种改善盐胁迫大麦根系液泡膜功能的机理

研究了0.1 mmol/L 腐胺 (Put) 和0.5 mmol/L 亚精胺 (Spd) 浸种对200 mmol/L NaCl胁迫下大麦(Hordeum vulgare L.)幼苗生长速率、干物质积累、离子分布、液泡膜蛋白结合多胺含量以及液泡膜膜脂组分与功能的影响.结果表明,Put和Spd浸种均可缓解盐胁迫对大麦幼苗的盐害,促进生长和干物质积累,降低大麦幼苗体内[Na+]/[K+].与盐处理的对照植株相比,Put和Spd浸种均可提高大麦幼苗根系液泡膜磷脂含量,降低糖脂结合半乳糖含量,而膜上非共价结合多胺含量Spd+PAx (一种未知多胺) 与 Put+Dap (二氨基丙烷)之比((Spd+PAx)/(Put+Dap))、共价和非共价结合多胺总量均上升.统计分析结果表明,液泡膜非共价结合多胺(Spd+PAx)/(Put+Dap)与H+-ATPase和H+-PPase活性呈显著正相关关系.     

氯化钠胁迫对嫁接黄瓜叶片多胺含量的影响

以日本耐盐品种‘帝王新土佐’南瓜为砧木,以’新泰密刺’黄瓜为接穗,在100 mmol·L^-1 NaCl胁迫下,对黄瓜嫁接和自根植株不同时期叶片中不同形态多胺含量的变化进行了研究.结果表明：NaCl胁迫下黄瓜嫁接植株游离态腐胺（Put）含量在胁迫2 d时与自根植株无显著差异,其余时间均显著高于自根植株;游离态亚精胺（Spd）和游离态精胺（Spm）含量在整个胁迫期间均显著高于自根植株;游离态多胺总量（PAs）在胁迫第4天出现峰值;嫁接植株游离态Put/PAs值在胁迫4 d时与自根植株无显著差异,其余胁迫时间均显著低于自根植株,而(Spd+Spm)/Put值在整个胁迫期间均显著高于自根植株;嫁接植株结合态和束缚态Put、Spd和Spm含量在整个胁迫期间均显著高于自根植株,结合态和束缚态PAs在胁迫第6天出现峰值;结合态多胺的Put/PAs值和(Spd+Spm)/Put值变化趋势与游离态多胺一致;嫁接植株束缚态Put/PAs值在胁迫6 d时与自根植株无显著差异,其余时间均显著低于自根植株,而(Spd+Spm)/Put值在整个胁迫期间均显著高于自根植株.表明黄瓜嫁接植株表现出较强的耐盐特征.     

低氧胁迫下24-表油菜素内酯对黄瓜幼苗叶片光合特性及多胺含量的影响

采用1/2 Hoagland营养液培养,研究了低氧胁迫下24-表油菜素内酯(EBR)对黄瓜幼苗叶片光合特性及多胺含量的影响.结果表明：低氧胁迫下黄瓜幼苗的净光合速率(P_n)、气孔导度(g_s)、蒸腾速率(T_r)、胞间CO₂浓度(C_i)显著下降,而叶绿素含量显著提高,幼苗生长受抑;低氧胁迫显著提高了黄瓜幼苗叶片的腐胺(Put)、亚精胺(Spd)、精胺(Spm)、多胺(PAs)含量和Put/PAs,但降低了(Spd+Spm) /Put.低氧胁迫下,外源EBR不仅显著提高了黄瓜幼苗的P_n、g_s、T_r及叶绿素含量,也显著提高了黄瓜幼苗叶片的游离态Spm、结合态Spd、Spm及束缚态Put、Spd、Spm含量,促进了PAs的进一步积累,且降低了Put/PAs,提高了(Spd+Spm)/Put．可见,外源EBR调节了黄瓜幼苗内源多胺含量及形态的变化,维持了较高的光合性能,促进了叶面积和干物质量的显著增加,缓解了低氧胁迫对黄瓜幼苗的伤害.     

Putrescine promotes changes in the endogenous polyamine levels and proteomic profiles to regulate organogenesis in <Emphasis Type="Italic">Cedrela fissilis</Emphasis> Vellozo (Meliaceae)

Improvements to in vitro organogenesis are essential for optimizing shoot development and understanding basic physiological processes. The addition of polyamines (PAs) to the culture medium has been used to modulate organogenesis in plants, and this work evaluated the effects of exogenous PAs on direct organogenesis from apical and cotyledonary nodal Cedrela fissilis explants as well as the effects of putrescine (Put) on endogenous PA levels and variations in protein abundance. The effects of exogenous Put, spermidine, and spermine at 0, 0.5, 1, 2.5, or 5 mM on shoot development were tested. The comparison of the tested PAs to the control treatment revealed that 2.5 mM Put significantly increased the length of shoots from cotyledonary nodal explants, which are more sensitive than apical nodal explants, and treatment with 2.5 mM Put significantly increased the endogenous total free-PA and free-Put levels in shoots compared with the control (no Put). A comparative proteomic analysis of shoots indicated that 2.5 mM Put significantly changed the abundance of proteins, primarily metabolic and cellular proteins associated with stress and energy processes such as cell division. These results show that Put functions in endogenous PA metabolism and alters protein abundance, thereby contributing to shoot development in C. fissilis.     

Role of polyamines in adventitious shoot morphogenesis from cotyledons of cucumber <Emphasis Type="Italic">in vitro</Emphasis>

The relationship between polyamines (PAs) metabolism and adventitious shoot morphogenesis from cotyledons of cucumber was investigated in vitro. The endogenous levels of free putrescine (Put) and spermidine (Spd) in the explants decreased sharply, whereas endogenous spermine (Spm) increased during adventitious shoot morphogenesis. The presence of 1–15 mM Put, 1–2 mM Spd, 0.05–1 mM Spm, 5–10 M aminoethoxyvinylglycine (AVG) or 5 M AVG together with 50 M 1-aminocyclopropane-1-carboxylic acid (ACC) in the regeneration medium could promote adventitious shoot formation. Conversely, 1–5 mM D-arginine (D-Arg) or 0.01–0.1 mM methylglyoxal bis-guganylhydrazone (MGBG) inhibited regeneration; and 0.005–0.05 mM ACC displayed little or no evident effects. The explants growing on medium containing 5 M AVG produced higher levels of free Put and Spm, and on medium containing 5 mM Put the explants responded similarly to the AVG-treated explants. However, the exogenous use of 1 mM D-Arg reduced the levels of Put, Spd and Spm, and 0.1 mM MGBG reduced the levels of free Spd and Spm. Moreover, although the explants cultured on medium containing Put and MGBG enhanced ethylene production, AVG and D-Arg inhibited ethylene biosynthesis. This study shows the PAs requirement for the formation of adventitious shoot from cotyledons of cucumber in vitro and the enhanced adventitious shoot morphogenesis may be associated with the elevated level of endogenous free Spm, albeit the promotive effect of PAs on adventitious shoot morphogenesis may not be related to ethylene metabolism.     

Polyamines as biomarkers for plant regeneration capacity: improvement of regeneration by modulation of polyamine metabolism in different genotypes of indica rice

The importance of cellular polyamine (PA) levels and the ratio of putrescine (Put) to spermidine (Spd) for plant regeneration ability via somatic embryogenesis in several commercially grown indica rice varieties is reported here. The genotypes namely NDR-624, IR-20, IR-36, BJ-1 (having Put:Spd ratio2.3) showed superior plant regeneration while KL, PB-1 and TN-1 (having Put:Spd ratio3.8) showed moderate plant regeneration ability. The genotypes namely HS, Bindli, DV-85, ACB-72, IR-64 and IR-72 (having Put:Spd ratio5.0) showed poor plant regeneration ability. In contrast KH-7 (Put:Spd ratio10.0) showed no response at all. Favorable modification of cellular PA titers and their Put:Spd ratio by the addition of exogenous PAs (Put, Spd) or their biosynthesis inhibitor, difluoromethylarginine (DFMA) led to the induction/promotion of plant regeneration in poorly responding genotypes. These results showed a close relationship between cellular PA levels and their Put:Spd ratio with in vitro morphogenetic capacity in indica rice and suggest that the cellular PAs and Put:Spd ratios are important determinants (biomarkers) of plant regeneration ability in indica rice, and the improvement/induction of plant regeneration in morphogenetically poor and recalcitrant species could be achieved by modulating PA metabolism.     

Reinitiation of cell division and polyamine and aromatic monoamine levels in alfalfa explants during the induction of somatic embryogenesis

The early cellular events in leaf explants of Medicago sativa L. cultured on somatic embryogenesis- and callogenesis-inducing media (EIM and CIM, respectively) were correlated with the endogenous contents of polyamines (PAs) and aromatic monoamines (AMs). On the second day of culture, replication of DNA occurred in epidermal and subepidermal cells on the edges of explants on EIM and was a prerequisite for proembryonal mass and, later, globular proembryo formation. In explants cultured on CIM, replication occurred at least one day later and in fewer cells, which were randomly spread all over the explant. Transition of leaf explant cells to rapidly dividing meristematic-like cells on EIM or to enlarged, highly vacuolated cells on CIM was observed. The increase in total PA levels in cultured explants was primarily a consequence of increases in putrescine (Put) and spermidine (Spd) contents and was much more pronounced on EIM than on CIM. High Spd levels were characteristic of meristematic cells and might be essential for the development of globular structures. The higher amount of insoluble PA conjugates was determined in explant cells on EIM, as compared with CIM. Proembryogenic cell mass formation was positively correlated with free tyramine and negatively correlated with free phenylethylamine contents.     

多胺及其合成抑制剂MGBC对稀脉萍成花反应的影响

腐胺、亚精胺和精胺对稀脉萍的成花均有一定的抑制作用,这种作用随多肢的浓度增加而增强。多胺合成抑制剂MGBG强烈抑制稀脉萍群体的增殖速率,并使稀脉萍群体在非诱导光周期下开花。这种由MGBG引起的增殖速率的降低及成花诱导作用均可被多胺逆转。稀脉萍成花诱导过程中,内源腐胺含量显著升高,亚精胺则下降。