Arabidopsis ADC genes involved in polyamine biosynthesis are essential for seed development

Arginine decarboxylase (ADC) is a rate-limiting enzyme that catalyzes the first step of polyamine (PA) biosynthesis in Arabidopsis thaliana. We generated a double mutant deficient in Arabidopsis two ADC genes (ADC1-/- ADC2-/-) and examined their roles in seed development. None of the F2 seedlings from crosses of adc1-1 and adc2-2 had the ADC1-/- ADC2-/- genotype. In addition, some abnormal seeds were observed among the ADC1+/- ADC2-/- and ADC1-/- ADC2+/- siliques. Viable offspring with the ADC1-/- ADC2-/- genotype could not be obtained from the ADC1+/- ADC2-/- and ADC1-/- ADC2+/- plants. These results indicate that AtADC genes are required for production of polyamines that are essential for normal seed development in Arabidopsis.     

Systematic analysis of rice (Oryza sativa) metabolic responses to herbivory

Plants defend against attack from herbivores by direct and indirect defence mechanisms mediated by the accumulation of phytoalexins and release of volatile signals, respectively. While the defensive arsenals of some plants, such as tobacco and Arabidopsis are well known, most of rice's (Oryza sativa) defence metabolites and their effectiveness against herbivores remain uncharacterized. Here, we used a non‐biassed metabolomics approach to identify many novel herbivory‐regulated metabolic signatures in rice. Most were up‐regulated by herbivore attack while only a few were suppressed. Two of the most prominent up‐regulated signatures were characterized as phenolamides (PAs), p‐coumaroylputrescine and feruloylputrescine. PAs accumulated in response to attack by both chewing insects, i.e. feeding of the lawn armyworm (Spodoptera mauritia) and the rice skipper (Parnara guttata) larvae, and the attack of the sucking insect, the brown planthopper (Nilaparvata lugens, BPH). In bioassays, BPH insects feeding on 15% sugar solution containing p‐coumaroylputrescine or feruloylputrescine, at concentrations similar to those elicited by heavy BPH attack in rice, had a higher mortality compared to those feeding on sugar diet alone. Our results highlight PAs as a rapidly expanding new group of plant defence metabolites that are elicited by herbivore attack, and deter herbivores in rice and other plants.     

Remodeling of tobacco thylakoids by over-expression of maize plastidial transglutaminase

Transglutaminases (TGases, EC 2.3.2.13) are intra- and extra-cellular enzymes that catalyze post-translational modification of proteins by establishing ?-(γ-glutamyl) links and covalent conjugation of polyamines. In chloroplast it is well established that TGases specifically polyaminylate the light-harvesting antenna of Photosystem (PS) II (LHCII, CP29, CP26, CP24) and therefore a role in photosynthesis has been hypothesised (Della Mea et al. [23] and refs therein). However, the role of TGases in chloroplast is not yet fully understood. Here we report the effect of the over-expression of maize (Zea mays) chloroplast TGase in tobacco (Nicotiana tabacum var. Petit Havana) chloroplasts. The transglutaminase activity in over-expressers was increased 4 times in comparison to the wild-type tobacco plants, which in turn increased the thylakoid associated polyamines about 90%. Functional comparison between Wt tobacco and tgz over-expressers is shown in terms of fast fluorescence induction kinetics, non-photochemical quenching of the singlet excited state of chlorophyll a and antenna heterogeneity of PSII. Both in vivo probing and electron microscopy studies verified thylakoid remodeling. PSII antenna heterogeneity in vivo changes in the over-expressers to a great extent, with an increase of the centers located in grana-appressed regions (PSIIα) at the expense of centers located mainly in stroma thylakoids (PSIIβ). A major increase in the granum size (i.e. increase of the number of stacked layers) with a concomitant decrease of stroma thylakoids is reported for the TGase over-expressers.     

Attitudes Toward Resolution of Human–Wildlife Conflict Among Forest-Dependent Agriculturalists Near Rajaji National Park,India

Understanding local attitudes towards human–wildlife conflict (HWC) is key to developing successful conflict mitigation strategies. In this paper, in-depth interview and questionnaire data about resolution of HWC in Uttarakhand, India are examined from both qualitative and quantitative approaches (n = 70). Responses are differentiated between and within three subgroups: gender, literacy status, and relative wealth. Overall, the plurality of respondents said that fencing is the best solution, that the Forest Department should take leadership, and that villagers would be willing to participate in a cooperative management institution. However, cooperative action was only actively supported by 27.4% of respondents, suggesting that comanagement of this protected area will require significant capacity building and trust building activities. Intragroup differences show that all three factors are significant, and underscore the importance of addressing gender differences in attitudes about HWC in particular. Women were less likely than men to support compensation, more likely to prefer that the village take leadership, and less willing to participate in a cooperative management institution. The study illustrates the value of mixed-method research, and suggests a number of specific entry points for action.

NaCl胁迫对白刺幼苗体内游离态亚精胺和精胺含量的影响

以西伯利亚白刺试管苗为材料,研究其在0、50和200 mmol·L-1NaCl胁迫及外源环己胺调控下,生物量和体内多胺含量的改变,以探讨多胺对盐生植物白刺在盐胁迫下的生理调节功能.结果显示:50 mmol·L-1NaCl更有利于西伯利亚白刺的生长,处理后30 d白刺根系和地上部干重分别比对照增加75%和95%,处理15 d生根率比对照提高44.42%.而0(对照)和200 mmol·L-1NaCl都显著抑制白刺试管苗的生根与生长;200 mmol·L-1NaCl处理45 d时地上部干重比对照显著降低11.71%.白刺叶片中的多胺(精胺和亚精胺)含量随着盐浓度的增加呈降低趋势,茎和根中的多胺含量在200 mmol·L-1NaCl处理后才显著受到抑制;同时施环己胺和200 mmol·L-1NaCl处理使白刺叶片中的亚精胺含量显著低于200 mmol·L-1NaCl处理组,且叶绿素合成减少,细胞质膜透性增高.研究表明,西伯利亚白刺叶片内保持合适的多胺含量可能是其适应盐渍环境的重要机制之一.     

Amelioration of salinity stress by exogenously applied spermidine or spermine in three varieties of indica rice differing in their level of salt tolerance

We present here the comparative protective potentiality of exogenously applied polyamines (PAs), namely spermidine (Spd) and spermine (Spm), in mitigating NaCl toxicity and inducing short-term salinity tolerance in three indica rice varieties, namely M-1-48 (salt-sensitive), Nonabokra (salt-tolerant) and Gobindobhog (highly sensitive). The retardation in root length or shoot length and toxic Na⁺ accumulation or K⁺ loss, the considerable increment in malondialdehyde/H₂O₂ accumulation or lipoxygenase activity, all of which were particularly noteworthy in M-1-48 and Gobindobhog during salinity stress, was appreciably reduced by co-treatment with Spd or Spm. Both the PAs also inhibited the extent of salt-induced protein carbonylation in all the varieties and enhanced protease activity, especially in Gobindobhog. The prevention of chlorophyll degradation was better with Spd in Nonabokra and Gobindobhog. While the salt-induced increase in anthocyanin or reducing sugar level was further prompted by Spd or Spm in all the varieties, the proline content was elevated by Spd particularly in Gobindobhog. During salinity stress, both the PAs were effective in lowering the putrescine accumulation in M-1-48 and Gobindobhog, and strikingly increasing the Spm level in all the varieties, the highest being in Gobindobhog. In addition, they enhanced the activity of peroxidases and compensated for the decreased catalase activity in all the varieties. Thus the two PAs could recuperate all the three varieties from salt-induced damages to different degrees. The salt injuries, encountered in M-1-48 and Gobindobhog, both of which showed greater susceptibility to salinity stress, were more pronouncedly alleviated and counteracted by the PAs, than the salt-tolerant Nonabokra. The reversal of inhibitory effect of salinity stress was conferred by preventing growth inhibition or various forms of cellular damages, maintaining proper K⁺/Na⁺ balance or triggering the level of osmolytes and activity of antioxidant enzymes. Our communication offers a referenced evidence for an understanding of the mechanism by which higher PAs relieve the damages particularly in salt-sensitive rice varieties.     

Putrescine and Silver Nitrate Influences Shoot Multiplication, In Vitro Flowering and Endogenous Titers of Polyamines in Cichorium intybus L. cv. Lucknow Local

Abstract The influence of putrescine (Put) and AgNO₃ on shoot multiplication, in vitro flowering and endogenous titers of polyamines in Cichorium intybus L. cv. Lucknow local was investigated. Exogenous administration of Put at a concentration of 40 mM resulted in maximum tissue response in terms of shoot numbers (34.6 ± 2.61) and shoot lengths (7.6 ± 0.57 cm) on MS media supplemented with 2-iP (2.0 mg L⁻¹) and GA₃ (0.5 mg L⁻¹) as observed on the 35^th day. Exogenous application of 40 μM AgNO₃ resulted in maximum shoot number (36.8 ± 2.63) and shoot lengths (7.9 ± 0.76 cm) on day 35 on the same media. Endogenous titers of conjugated spermidine decreased sharply from day 7–21, whereas endogenous conjugated spermine levels peaked on day 28 (1265 ± 94.9 nmoles g⁻¹ FW), after treatment with 40 mM Put. Whereas, AgNO₃ (40 μM) fed samples resulted in higher titers of endogenous conjugated spermine (1405 ± 105.6 nmoles g⁻¹ FW, 3.62 fold over control) on day 14. All other treatments showed decreasing endogenous levels during the whole culture period. Both Put (40 mM) and AgNO₃ (40 μM) resulted in floral initiation and floral development on day 28 and 14 (3.76 ± 0.16, 4.2 ± 0.21 flowers per shoot apices), respectively. To investigate the role of Put (40 mM) and AgNO₃ (40 μM) on morphogenetic response and endogenous conjugated polyamine titers in shoots of chicory, polyamine inhibitors (DFMA and DFMO) were used. The morphogenetic response and the endogenous conjugated pool of polyamines were diminished in DFMA and DFMO treatments, but could be restored by addition of Put (40 mM) and AgNO₃ (40 μM). Under exogenous Put feeding, ethylene production was reduced in shoot cultures of chicory. This study shows for the first time the influence of polyamines on multiple shoot initiation from axillary buds of C. intybus L. cv. Lucknow local and also indicates the promotive effect of Put and AgNO₃ on autoregulation of polyamine biosynthesis, thereby regulating in vitro flowering, the endogenous pool of polyamines and shoot multiplication. Received 12 November 1999; accepted 11 February 2000     

靶向多胺代谢调控网络的小分子药物及抗肿瘤应用

多胺代谢调控网络包括多胺的生物合成、分解代谢和膜转运,作为生物体重要而复杂的生化单元,广泛参与机体细胞的生长、增殖、凋亡和基因表达等活动。多胺代谢调控网络的失衡与多种疾病相关,例如肿瘤、炎症和心血管疾病等。2018年全球癌症统计数据预计,癌症将成为21世纪几乎每个国家或地区人口死亡的主要原因。因此,癌症的预防和治疗将越来越重要。鉴于多胺与肿瘤的发生发展密切相关,本文围绕多胺代谢调控网络,总结了该调控网络作为抗肿瘤治疗靶位的研究现状,同时列举几种代谢酶和转运蛋白质的小分子调节剂,并阐述其靶点作用方式和在肿瘤预防与治疗方面的应用,以期能为靶向多胺代谢调控网络的药物研发以及相关疾病的治疗提供参考。     

Effects of polyamines on the functionality of photosynthetic membrane in vivo and in vitro

The three major polyamines are normally found in chloroplasts of higher plants and are implicated in plant growth and stress response. We have recently shown that putrescine can increase light energy utilization through stimulation of photophosphorylation [Ioannidis et al., (2006) BBA-Bioenergetics, 1757, 821-828]. We are now to compare the role of the three major polyamines in terms of chloroplast bioenergetics. There is a different mode of action between the diamine putrescine and the higher polyamines (spermidine and spermine). Putrescine is an efficient stimulator of ATP synthesis, better than spermidine and spermine in terms of maximal % stimulation. On the other hand, spermidine and spermine are efficient stimulators of non-photochemical quenching. Spermidine and spermine at high concentrations are efficient uncouplers of photophosphorylation. In addition, the higher the polycationic character of the amine being used, the higher was the effectiveness in PSII efficiency restoration, as well as stacking of low salt thylakoids. Spermine with 50 μM increase F_V as efficiently as 100 μM of spermidine or 1000 μM of putrescine or 1000 μM of Mg²⁺. It is also demonstrated that the increase in F_V derives mainly from the contribution of PSIIα centers. These results underline the importance of chloroplastic polyamines in the functionality of the photosynthetic membrane.