Properties of the System for the Mixed Function Oxidation of Kaurene and Kaurene Derivatives in Microsomes of the Immature Seed of Marah macrocarpus: Cofactor Requirements

The rates of oxidation of ent-kaur-16-ene to ent-kaur-16-en-19-ol, ent-kaur-16-en-19-al, ent-kaur-16-en-19-oic acid, and ent-kaur-16-en-7alpha-ol-19-oic acid are maximal in microsomes prepared from the endosperm of immature Marah macrocarpus seeds in which the cotyledons are approximately one-half the overall length of the seed. The supernatant fraction remaining from the preparation of the microsomes contains factors which stimulate the rates of oxidation catalyzed by the microsomes. Added TPNH is more effective than added DPNH in meeting the requirement for reduced pyridine nucleotide. A mixture of DPNH, ATP, and TPN(+) is much more effective than DPNH alone. Experiments with 2,4-dinitrophenol as a selective inhibitor indicate that the ATP-stimulated synthesis of TPNH which occurs in these microsomes in the presence of this mixture of coenzymes provide TPNH for use in the mixed function oxidations. Relatively low concentrations of DPNH and TPNH together are much more effective than either alone at equivalent concentration. This is consistent with the involvement of two pathways of electron transfer associated with the mixed function oxidations, one of which preferentially utilizes TPNH and the other favoring DPNH. FAD added to microsomes at an optimal concentration of about 10 mum in the presence of TPNH stimulates the rate of the oxidations; higher concentrations are inhibitory. FMN by itself does not produce this stimulation. However, FMN and FAD added together at low concentrations (0.5 mum each) have approximately the same effectiveness as FAD alone at 10 mum. This suggests a role for both flavin nucleotides in the normal electron transfer pathways associated with these oxidations. Some of the stimulatory properties of the supernatant fraction may be accounted for by its content of reduced pyridine nucleotides, FAD, and FMN; the concentrations of FAD and FMN were determined to be 1.1 mum and 0.4 mum, respectively. However, the effects of the supernatant fraction are not completely explained by its content of these coenzymes since other experiments indicate the presence of a heat-labile, nondialyzable stimulatory factor(s) in the supernatant fraction in addition to heat-stable, dialyzable fractors.     

Properties of the System for the Mixed Function Oxidation of Kaurene and Kaurene Derivatives in Microsomes of the Immature Seed of Marah macrocarpus: Electron Transfer Components

Cytochrome P-450 and cytochrome b₅ at levels of approximately 0.10 and 0.60 nanomole per milligram of microsomal protein were detected by spectral measurements in microsomes prepared from endosperm tissue of immature Marah macrocarpus seeds. TPNH-cytochrome c reductase, DPNH-cytochrome c reductase, andDPNH-cytochrome b₅ reductase activities were also present in these microsomes at levels of approximately 0.060, 0.22, and 0.52 unit per milligram of microsomal protein, respectively. (One unit of reductase is the amount of enzyme catalyzing the reduction of 1 micromole of electron acceptor per minute.) Treatments of microsomes with steapsin or trypsin were not effective in solubilizing any of these electron transport components in detectable form. However, treatment of a microsomal suspension in 25% glycerol with 1% sodium deoxycholate led to the release of about 60% of the protein and each of the above hemoproteins and electron transfer activities to the fraction which was not pelleted after centrifugation for 2 hours at 105,000g. Some ent-kaur-16-ene oxidase activity could be detected in the solubilized fraction after removal of the detergent. Cytochrome b₅ and DPNH-cytochrome b₅ reductase activity were largely separated from one another and from an overlapping mixture of TPNH-cytochrome c reductase and DPNH-cytochrome c reductase when the sodium deoxycholate-solubilized fraction was chromatographed on a DEAE-cellulose column. No cytochrome P-450 or cytochrome P-420 was detected in the column fractions and no ent-kaur-16-ene oxidase activity was detected when the column fractions were tested singly or in combination.     

Biosynthetic Origin of Gibberellins A(3) and A(7) in Cell-Free Preparations from Seeds of Marah macrocarpus and Malus domestica

Cell-free preparations from seeds of Marah macrocarpus L. and Malus domestica L. catalyzed the conversion of gibberellin A₉ (GA₉) and 2,3-dehydroGA₉ to GA₇; GA₉ was also metabolized to GA₄ in a branch pathway. The preparation from Marah seeds also metabolized GA₅ to GA₃ in high yield; GA₆ was a minor product and was not metabolized to GA₃. Using substrates stereospecifically labeled with deuterium, it was shown that the metabolism of GA₅ to GA₃ and of 2,3-dehydroGA₉ to GA₇ occurs with the loss of the 1β-hydrogen. In cultures of Gibberella fujikuroi, mutant B1-41a, [1β,2β-²H₂]GA₄, was metabolized to [1,2-²H₂]GA₃ with the loss of the 1α- and 2α-hydrogens. These results provide further evidence that the biosynthetic origin of GA₃ and GA₇ in higher plants is different from that in the fungus Gibberella fujikuroi.     

Growth Promoting Activities of Sclerotinin A and Its Analogs

(±)-Sclerotinin A (Scl. A) showed the same promoting activity as natural (+)-Scl. A, which was isolated as a growth promoting substance of rice seedlings from Sclerotinia sclerotiorum along with sclerin. By the combined use of Scl. A and gibberellin, the synergistic effect on the growth of rice seedlings was noticed as in the case of sclerin. Correlation between the chemical structure and biological activity of Scl. A derivatives and analogs was discussed.     

Effects of a Plant-Growth Regulator, Prohexadione, on the Biosynthesis of Gibberellins in Cell-Free Systems Derived from Immature Seeds

Inhibition of the biosynthesis of gibberellins by prohexadione,3,5-dioxo-4-propionylcyclo-hexanecarboxylic acid, was studiedwith cell-free systems derived from immature seeds of Cucur-bitamaxima, Phaseolus vulgaris and Pisum sativum. Prohexadione,at a concentration of 10–⁴ M, inhibited C-7 oxidationof GA₁₂-aldehyde, C-20 oxidation of GA₁₅, conversion of C₂₀-gib-berellinsto C₁₉-gibberellins, 3ß-hydroxylation, 2,3-dehydrogenationof GA²⁰, 2,3-epoxidation of GA₅ and 2ß-hydroxylationof GA₉ and GA₂₀. The 3ß-hydroxylase activity appearedto be more sensitive to prohexadione than were the C-20 oxygenaseand the 2ß-hydroxylase activities. The conversionof mevalonic acid to GA₁₂-aldehyde and the 13-hydroxylationof GA₁₂ were not affected by prohexadione at a concentrationof 3 ? 10^–4 M. All of the steps inhibited by prohexadioneare oxidation steps catalyzed by soluble enzymes that require2-oxoglutarate, Fe²⁺ and oxygen, and all of them occur distalto the synthesis of GA₁₂-aldehyde in the biosynthesis of gibberellins. (Received April 4, 1990; Accepted September 14, 1990)     

Metabolism of Gibberellins in Cell-Free Extracts of Anthers from Normal and Dwarf Rice

Cell-free extracts were prepared from anthers of normal anddwarf rice (Oryza sativa L.), and the metabolism of radioisotope-labeledgibberellins in the extracts was analyzed by HPLC and gas chromatography-massspectrometry (GC/MS). GA₁₂ was converted to GA₁₅ and GA₃₄ inthe extracts. GA₂₀ was converted to GA¹, GA₈ and GA₂₉, but GA₉was converted only to GA₃₄. The extracts of the dwarf cultivar,Waito-C (dy mutant), showed the same 3ß-hydroxylationactivity as did those of the normal cultivar, Nihonbare, indicatingthat the dy gene is not expressed in the anthers. These resultssuggest that the regulation of the biosynthesis of gibberellinsin rice is organ-specific. (Received November 9, 1989; Accepted January 10, 1990)     

77种植物提取物对植物病原菌的生长抑制作用

以稻瘟病菌(Pyricularia grisea),水稻恶苗病菌(Fusarium moniliforme),玉米大斑病菌(Exserohilum turci-cum)等12种植物病原菌为供试菌种,采用生长速率法对77种植物的95%乙醇提取物在200μg/mL下进行室内抑菌试验。结果表明有15种植物提取物对植物病原菌有抑制作用,其中华山姜、硬骨藤、龙舌兰、红蒜、大花哥纳香、海南草珊瑚对至少一种菌的抑制率在50%以上,版纳青梅、大果巴戟、华山姜等8种植物提取物对至少三种病原菌均有不同程度的抑制作用。红蒜提取物对百合炭疽病菌、海南草珊瑚提取物对番茄灰霉病菌,以及龙舌兰提取物对玉米大斑病菌的抑制率分别为61.4%、70.7%、76.6%,与阳性对照抑制率相比效果明显。     

Effects of Vitamin A and Its Analogs on Nonenzymatic Lipid Peroxidation in Rat Brain Mitochondria

Vitamin A (retinol) and some of its analogs exhibited varying degrees of inhibition on induced iron and ascorbic acid lipid peroxidation of rat brain mitochondria. Malonyldialdehyde production was used as an index of the extent of in vitro lipid peroxidation. The fat-soluble vitamins retinol, retinol acetate, retinoic acid, retinol palmitate, and retinal at concentrations between 0.1 and 10.0 mmol/L inhibited brain lipid peroxidation. Retinol and retinol acetate were the most effective inhibitors. It is concluded from this study that retinol and its analogs can be considered as potential antioxidant factors, more potent than some of the well-known antioxidants such as alpha-tocopherol and butylated hydroxytoluene.     

ent-Kaurene Biosynthesis in Cell-Free Extracts of Excised Parts of Tall and Dwarf Pea Seedlings

Investigations on the sites of ent-kaur-16-ene (ent-kaurene) biosynthesis were conducted with cell-free extracts from several excised parts of 10-, 13-, and 16-d-old tall and dwarf pea (Pisum sativum L.) seedlings. [¹⁴C]Mevalonic acid was incorporated into ent-kaurene in cell-free extracts from young developing leaves and elongating internodes of tall (`Alaska') and dwarf (`Progress No.9') pea seedlings at all three stages of development. ent-Kaurene biosynthesis also occurred readily in cell-free extracts from shoot tips, petioles, and stipules near the young elongating internodes. The ent-kaurene-synthesizing activity found in young developing tissues declined as tissues matured. Little or no activity was detectable in enzyme extracts from cotyledons and root tips at different stages. In light grown tall pea internodes ent-kaurene-synthesizing activity was low as they began to elongate, reached a maximum when the internodes reached about 2 cm in length and declined as they matured. Activity in extracts of dwarf shoot tips and internodes was generally lower than in equivalent tall plants, but the activity in dwarf leaves and stipules was somewhat higher than in tall plants. With the exception of root tips, there is a strong correlation between growth potential of a tissue and the rate of ent-kaurene biosynthesis in extracts from that tissue.     

天然植物提取物对肉鸭生长性能和屠宰性能的影响

采用单因子试验设计,研究了天然植物提取物对生长后期肉鸭生长性能和屠宰性能的影响.选择健康、体重相近(659.05±8.04 g)的14日龄天府肉鸭120只(公母各半),随机分为5个处理(4个试验组和1个对照组),每个处理4个重复,每个重复6只鸭(公母各半).基础日粮中分别添加37.5、75、150和225 mg/kg植物提取物即为相应的试验日粮,对照组饲喂基础日粮.试验期28 d.结果表明:①除225 mg/kg组降低采食量外,其余各组对采食量的影响不显著;②日粮中添加不同剂量植物提取物均提高了肉鸭试验各期的生长速度和饲料转化效率,与对照组相比,37.5,75,150和225 mg/kg植物提取物分别提高全期ADG 6.30%(P<0.05),7.07%(P<0.05),9.62%(P<0.01)和5.28%(P<0.05),分别降低全期F/G 4.60%(P<0.05),10.73%(P<0.01),8.81%(P<0.01)和14.56%(P<0.01).回归分析表明,全期ADG随植物提取物添加剂量呈显著二次曲线(P<0.05)上升,当添加剂量为132.75 mg/kg时,试验全期增重速度最大,达86.8 g/d;全期F/G随添加剂量增加呈极显著二次曲线下降(P<0.01);③植物提取物显著改善肉鸭的屠宰性能,显著提高瘦肉率(P<0.05),极显著降低腹脂沉积(P<0.01),且随添加剂量增加呈极显著二次曲线关系变化(P<0.001);④植物提取物在生长后期肉鸭日粮中的适宜添加剂量为150 mg/ks.结果提示,植物提取物可通过提高饲料利用效率显著改善生长后期肉鸭的生长性能和屠宰性能,且存在剂量效应.     

Effect of Monuron and of Related Compounds on the Growth of Seedling Peas

Shoots of peas grown in phosphate buffer were no more sensitiveto monuron applied continuously than were roots. They were,however, more readily damaged than roots when high monuron concentrationswere applied for short periods of time. Neither root nor shootwas more sensitive to monuron in light than in darkness. Thedamage caused by a high monuron concentration was far greaterat times, or under conditions, of high metabolic activity. Thetoxicities of alkyl or aryl substituted ureas and thioureaswere closely related to their solubilities in water. In viewof this similarity with physical poisons and the probable sitesof their main toxic actions, it is considered more likely thatthey disrupt an integrating surface than that they act uponspecific active groups of enzymes.