Synthesis of phenylalanine ammonia-lyase in anthocyanin-containing and anthocyanin-free callus cells of Daucus carota L.

When callus cells of Daucus carota are grown on a medium containing gibberellic acid (GA₃) in a physiological concentration of 3x10^-6 M the cells cease to accumulate anthocyanins. This anthocyanin-free cell line has a very low activity of phenylalanine ammonia-lyase. After density labelling with D₂O an intensive de novo synthesis of the phenylalanine ammonia-lyase (E.C. 4.3.1.5; PAL) in the anthocyanin-containing cells does occur. 58% of the C-bound H-atoms are replaced by deuterium. The anthocyanin-free cells show only a very low enzyme synthesis which is difficult to detect with density labelling experiments. To ascertain that de novo synthesis occurs in the anthocyanin-free cells, the incorporation of ¹⁴C-labelled amino acids into the partially purified enzyme protein was measured after separation of the protein a) in CsCl gradients and b) on polyacrylamide gels. In both cases the enzyme bears ¹⁴C-label. These results suggest that in the anthocyanin-free cells de novo synthesis of PAL is still occuring but the synthesis is reduced in comparison to the anthocyanin-containing cells.Abbreviations GA₃ gibberellic acid - PAL phenylalanine ammonia-lyase (E.C.4.3.1.5) - DCb anthocyanin-containing cells - DCw anthocyanin-free cells     

生长调节剂对离体银杏叶苯丙氨酸解氨酶活性的影响

用6种生长调节剂诱导离体银杏(Ginkgo biloba Linn.)叶苯丙氨酸解氨酶(PAL)活性,结果表明:300mg·L-1ETP诱导作用最强,其动态诱导在处理4 h后最高;40 mg·L-1 2,4-D诱导效果最强,并有2个明显的诱导高峰;200mg·L-1CCC诱导作用最强,在处理8 h后出现诱导高峰;除100mg·L-1IAA处理组的酶活性比对照低外,其他浓度的IAA处理均比对照高,诱导作用最强的是70 mg·L-1IAA处理;除了100mg·L-1ABA处理使酶活性略有降低外,其他浓度的ABA处理都能诱导酶活性升高,以75 mg·L-1ABA诱导能力最强,在处理4 h后酶活性最大;4个浓度的GA处理中,仅75 mg·L-1GA处理组的酶活性高于对照,处理8 h后酶活性达到最大.结果说明诱导效果从高至低依次为:乙烯利(ETP)、2,4-D、矮壮素(CCC)、吲哚乙酸(IAA)、脱落酸(ABA)、赤霉素(GA).     

甘蓝幼苗受桃蚜危害后叶片中部分酶活性的变化

对桃蚜（Myzus persicae Sulzer）危害后甘蓝（Brassica oleracea L.）幼苗叶中多酚氧化酶（PPO）、苯丙氨酸解氨酶（PAL）、过氧化物酶（POD）活性及同工酶进行了研究。结果表明,随桃蚜危害时间延长,处理叶片PPO、PAL和POD的活性与对照相比均表现出升高的趋势,方差分析表明与对照之间差异显著。同工酶电泳结果表明,POD同工酶的部分谱带随危害时间而发生变化,但EST同工酶在接虫前后则无明显的变化。     

红豆杉细胞周期时相与紫杉醇诱导合成的关系初探

利用植物细胞培养生产药用次生代谢产物 ,普遍存在着次生代谢物产量不稳定的问题。次生代谢物是一大类无明显生理功能或者非生长发育所必需的小分子有机化合物 ,其生物合成与积累一般发生在植物某些器官和组织发育的某个时期。因此 ,植物细胞的生长发育状态与次生代谢物产量有密切的关系。细胞周期时相在很大程度上反映细胞本身所处的状态 ,对细胞周期时相进行调控可能对次生代谢物产量具有重要影响 [1 ] 。诱导作用是提高植物次生代谢物产量的有效方法之一 [2 ,3]。我们以 2种经典同步化方法 ,建立红豆杉悬浮细胞初始周期时相模型 ,对不同…     

棉花抗蚜性与苯丙氨酸解氨酶活性的关系

为探讨苯丙氨酸解氨酶活性与棉花抗蚜性的关系,在室内人工接蚜危害和田间自然蚜群2种条件下,测定不同抗蚜性棉花品种受棉蚜Aphis gossypii Glover危害后叶片的PAL活性。结果表明:未受棉蚜危害时,抗、感品种PAL活性无明显差异;棉蚜危害胁迫棉花PAL活性升高,抗蚜品种受到棉蚜危害诱导所产生的PAL活性远比感蚜品种高。田间有蚜株率与棉花叶片苯丙氨酸解氨酶活性的呈显著的负相关关系。研究说明PAL酶活性对棉花的抗蚜性有一定的影响。     

Control of phenylalanine ammonia-lyase gene promoters from pea by UV radiation

The gene fusion system was used to study UV light-control of PS PAL1 and PS PAL2 genes encoding phenylalanine ammonia-lyase of pea. The induction of pea PAL promoters was analysed in transgenic tobacco plants. Binary plasmids (derivatives of pBI101.2 vector) containing 5′ regulatory fragments of PS PAL1 and PS PAL2 linked to reporter genes (GUS,LUC) were constructed. The analyses were performed with the use of single constructs (containing one variant of PS PAL promoter and one reporter gene) and dual constructs (containing both PS PAL1 and PS PAL2 promoters connected with different reporter genes). The use of dual constructs enabled the evaluation of both PS PAL promoters activity in the same plant. The analyses of in vitro grown plants have shown that both PAL promoters are strongly induced in leaves subjected to UV radiation. In some cases, the UV-stimulated expression exceeded the exposed areas. This phenomenon was observed more often in the leaves of plants containing the PS PAL1::GUS than PS PAL2::GUS construct. Removal of boxes 2, 4, 5 from PS PAL1 promoter and deletion of its 5′ end region (-339 to -1394) decreases the level of gene expression but does not eliminate its responsiveness to UV.     

Responses of anthocyanin-producing and non-producing cells of Glehnia littoralis to radical generators.

The responses of anthocyanin-producing (violet) and non-producing (white) cells of Glehnia littoralis to radical generators were compared. Cell growth, anthocyanin content, phenylalanine ammonia-lyase (PAL) activity and furanocoumarin production were determined after treatment with H(2)O(2), 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH), X-ray and yeast extract, independently. AAPH and H(2)O(2) repressed the growth of both violet and white cells, but violet cells grew better than white cells. On the other hand, the anthocyanin content in violet cells decreased. Neither X-ray nor yeast extract affected cell growth or pigment production. Treatment with H(2)O(2), yeast extract, and X-ray, but not AAPH, induced PAL activity and furanocoumarin production in white cell cultures, whereas violet cell cultures did not produce furanocoumarin following any of the treatment employed.     

Acibenzolar-S-methyl induced resistance to <Emphasis Type="Italic">Phytophthora capsici</Emphasis> in pepper leaves

The leaves of pepper (Capsicum anuum L.) were inoculated with Phytophthora capsici Leonian 3 d after treatment with acibenzolar-S-methylbenzo [1,2,3]thiadiazole-7-carbothioic acid-S-methyl ester (ASM) and resistance to Phytophthora blight disease was investigated. Results showed that P. capsici was significantly inhibited by ASM treatment by up to 45 % in planta. The pepper plants responded to ASM treatments by rapid and transient induction of L-phenylalanine ammonia-lyase (PAL), increase in total phenol content and activities of chitinase and β-1,3-glucanase. No significant increases in enzyme activities were observed in water-treated control plants compared with the ASM-treated plants. Therefore it may be suggested that ASM induces defense-related enzymes, PAL activity, PR proteins and phenol accumulation in ASM-treated plants and contribute to enhance resistance against P. capsici.     

The Arabidopsis phenylalanine ammonia lyase gene family: kinetic characterization of the four PAL isoforms

In Arabidopsis thaliana, four genes have been annotated as provisionally encoding PAL. In this study, recombinant native AtPAL1, 2, and 4 were demonstrated to be catalytically competent for l-phenylalanine deamination, whereas AtPAL3, obtained as a N-terminal His-tagged protein, was of very low activity and only detectable at high substrate concentrations. All four PALs displayed similar pH optima, but not temperature optima; AtPAL3 had a lower temperature optimum than the other three isoforms. AtPAL1, 2 and 4 had similar K(m) values (64-71 microM) for l-Phe, with AtPAL2 apparently being slightly more catalytically efficacious due to decreased K(m) and higher k(cat) values, relative to the others. As anticipated, PAL activities with l-tyrosine were either low (AtPAL1, 2, and 4) or undetectable (AtPAL3), thereby establishing that l-Phe is the true physiological substrate. This detailed knowledge of the kinetic and functional properties of the various PAL isoforms now provides the necessary biochemical foundation required for the systematic investigation and dissection of the organization of the PAL metabolic network/gene circuitry involved in numerous aspects of phenylpropanoid metabolism in A. thaliana spanning various cell types, tissues and organs.