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鱼腥藻SP.595液泡化原生质球诱导条件的研究 总被引:1,自引:0,他引:1
用浓度为0.15mol/L的KNO3、KCl、K2SO4、KH2PO4、K2HPO4、NaNO3、NaCl、Na2SO4、NaH2PO4、Na2HPO4、(NH4)2SO4、MgSO4、CaC12等单盐分别配合0.1%的溶菌酶处理鱼腥藻sp.595(Anabaenasp.595)。经4h,KH2PO4、K2SO4、Na2SO4、NaH2PO4、(NH4)2SO4、MgSO4、CaC12能诱导形成原生质球,但液泡化原生质球极少,而KNO3、NaNO3、NaC1诱导形成少量液泡化原生质球。用相近浓度的双盐,即KNO3和NaC1、KNO3和(NH4)2SO4、NaC1和(NH4)2SO4分别配合0.1%的溶菌酶处理,诱导效果亦不佳。用相近浓度的三盐NaC1、KNO3和(NH4)2SO4及五种盐NaC1、KNO3、(NH4)2SO4、Na2HPO4和KH2PO4分别配合0.1%的溶菌酶处理,诱导原生质球和液泡化原生质球的效果明显提高,液泡化原生质球比例达到66.90%—79.97%。 相似文献
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杂色云芝产漆酶的发酵条件研究* 总被引:3,自引:0,他引:3
本文对杂色云芝(Coriolus versicolor)产漆酶的发酵条件作了研究。结果表明摇瓶实验产漆酶(Laccase)的最佳培养基成分为:可溶性淀粉 2g/L, NH4Cl 24mmol/L, 微量元素混合液 7ml/L, pH3.0柠檬酸—Na2HPO4缓冲溶液 0.01mol/L, KH2PO4 1.4×10-2 mol/L, MgSO4·7H2O 2.03×10-3mol/L, CaCl2·2H2O 6.8×10-4 mol/L, VB1 2.97×10-6 mol/L, 吐温80 4.0g/L, 愈创木酚0.01mmol/L, CuSO4 ·5H2O 0.005mmol/L,最佳发酵条件为培养基初始pH3.0, 菌体生长6d,培养基装量为250ml三角瓶中25ml培养液,25℃条件下振荡培养(150r/min)9d。 相似文献
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采用响应面法对解淀粉芽孢杆菌C101菌株产芽孢发酵培养基进行了优化。利用Plackett-Burman试验设计筛选出影响产孢的3个主要因素:MnSO4、KH2PO4和(NH4)2SO4。在此基础上运用最陡爬坡路径法逼近最大响应值区域,最后利用响应面分析法确定主要因子之间的交互作用及最佳条件。结果表明,蔗糖20 g/L,尿素4.0 g/L,豆粕4.0 g/L,KNO3 2.0 g/L,Na2HPO4 2.4 g/L,KH2PO4 0.52 g/L,(NH4)2SO4 0.55 g/L,NaCl 1.0 g/L,MgSO4·7H2O 0.50 g/L,FeSO4 0.005 0 g/L,MnSO4 0.005 4 g/L,C101最大理论芽孢含量为14.67×108个/mL。经3次平行试验验证,实际平均芽孢含量与预测芽孢含量相近,比之前的芽孢含量提高了188%。 相似文献
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考察了在大肠杆菌中自诱导表达人胰高血糖素样肽-1突变体融合蛋白的可行性,并对自诱导培养条件及培养基成分进行优化,以提高蛋白产量。实验结果表明,最优培养基成分为蛋白胨19.17g/L,酵母膏9.59g/L,Na2HPO45.72g/L,KH2PO45.48g/L,(NH4)2SO42.66g/L,NaCl3.33g/L,甘油2%(V/V),葡萄糖0.68g/L,乳糖6.33g/L,MgSO40.24g/L。在温度33°C、接种量1%、pH7、装瓶量20mL/100mL培养条件下,用该最优培养基自诱导表达人胰高血糖素样肽-1突变体融合蛋白的产量可达348.6mg/L。 相似文献
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为了利用大肠杆菌高效生产重组磷脂酶,克隆了液化沙雷氏菌磷脂酶A1的编码基因pla,分别使用pET-28a(+)和pET-20b(+)载体,实现了磷脂酶A1在大肠杆菌BL21(DE3)中的功能表达.重组菌利用载体pET-28a(+)在原始信号肽的介导下胞外PLA1酶活达40.8 U/mL,占总酶活的91%.重组菌转接至优化后的发酵诱导培养基:蛋白胨10 g/L,酵母粉5g/L,葡萄糖0.8 g/L,乳糖5 g/L,25 mmol/L Na2HPO4,25 mmol/L KH2PO4和1 mmol/L MgSO4;菌体生长6h后,添加7.5 g/L的甘氨酸,37℃恒温发酵24 h,重组菌胞外PLA1酶活达到128.7 U/mL. 相似文献
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球衣菌合成聚羟基烷酸(PHA)的发酵研究 总被引:1,自引:0,他引:1
研究了球衣菌 (Sphaerotilussp.)W991 3 6合成聚羟基烷酸 (PHA)的培养基配方及发酵条件。结果表明 ,W991 3 6适宜发酵培养基配方为 :葡萄糖 1 0 2 5g/L ,蛋白胨 2 6 3g/L ,MgSO4·7H2 O 0 1 7g/L ,CaCl2 0 0 5g/L ,NaH2 PO4·2H2 O 0 0 2g/L ,K2 HPO40 0 4g/L ,KH2 PO40 0 3g/L ;最佳接种量为 0 1 3 8g (干 ) / 1 0 0mL ,培养基适宜初 相似文献
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以产琥珀酸放线杆菌Actinobacillus succinogenes NJ113 为出发菌株,针对该菌株筛选出含有关键生长因子的化学合成培养基,其关键因子为谷氨酸(Glu)、蛋氨酸(Met)和生物素(VH)和烟酸(VPP)。结合原发酵培养基中的磷酸缓冲盐成分,最终得到的化学合成培养基配方(g/L): CH3COONa 1.36,NaCl 1.0,MgCl2 0.2,CaCl2 0.2,Na2HPO4 0.31,NaH2PO4 1.6, KH2PO4 3,NH4HCO3 1.57,Glu 0.87,Met 0.11,VH 0.010,VPP 0.025。在3 L发酵罐上进行验证实验,50 g/L初始葡萄糖发酵70 h,丁二酸的质量浓度为45.2 g/L,丁二酸收率达到90.4%。与之前的半合成培养基发酵制备丁二酸相比,丁二酸的收率提高了25.2%,副产物也有很大幅度的减少。 相似文献
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Kiyoon Kang Kyungjin Lee Sangkyu Park Sungbeom Lee Young Soon Kim Kyoungwhan Back 《Journal of Plant Biology》2010,53(4):291-296
Protoporphyrin IX is a photosensitizer and a causative agent of rice membrane lipid peroxidation in plant cells. Protoporphyrinogen
IX oxidase (PPO) is the molecular target of PPO-inhibiting herbicides, which trigger a massive increase in protoporphyrin
IX. Thus, any possible method to decrease the levels of protoporphyrin IX upon challenge with PPO-inhibiting herbicides could
be employed to generate plants resistant to such herbicides. We generated transgenic rice plants overexpressing rice ferrochelatase
isogenes encoding ferrochelatase enzymes, which convert protoporphyrin IX into protoheme, to see whether the transgenic plants
have phenotypes resistant to PPO-inhibiting herbicides. The resulting transgenic rice plants were all susceptible to oxyfluorfen
(a diphenyl-ether-type PPO-inhibiting herbicide), as judged by cellular damage with respect to cellular leakage, chlorophyll
loss, and lipid peroxidation. In particular, the transgenic plants expressing rice ferrochelatase II without its plastid targeting
sequence showed higher transgene expression and oxyfluorfen susceptibility than lines expressing the intact ferrochelatase
II. Possible susceptibility mechanisms to oxyfluorfen herbicide in the transgenic rice plants are discussed. 相似文献
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Low temperature can greatly restrict the growth and development of rice. The rice seedlings show growth retardation, lamina wrap, and part of blade even died under the condition of low temperature. In order to get more information about cold stress responses in rice, two dimensional electrophoresis and bioinformatics analysis of mass spectrometry were used to preliminary survey the cold tolerance of cold sensitive line 9311 and cold resistance variety Fujisaka 5 under cold stress. Two dimensional electrophoresis maps of 9311 and Fujisaka 5 were established under cold treatment. With analysis of bioinformation, the proteins were found involve in many aspects of rice development. The largest category of proteins is functioning on metabolism. By comparing the proteins from the two varieties, it can be found that most proteins from 9311 were down-regulated and were up-regulated in Fujisaka 5. The results showed that the membrane composition and structure were damaged, metabolism changed dramatically and rice defense system was activated under the cold stimulation. Fifty-nine proteins related to the resistance of cold stress were identified in our study, and we have investigated and classified all of their biological functions. The importance of our study are providing some conduct for the research of rice resistant to cold stress, supporting auxiliary technique for rice varieties and widening the search field of cold tolerance in plants. 相似文献
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OPT8(511) was confirmed to be strongly associated with cold sensitivity of rice by random amplified polymorphic DNA (RAPD) analysis for the cold tolerance with 94 F2 population crossed with 'Dular' (cold sensitive cultivar) and 'Toyohatamochi' (cold resistant cultivar). A DNA marker from the RAPD fragment, OPT8(511), has been cloned with genomic DNA from rice cultivar ('Dular') and the nucleotide sequence has been determined. The nucleotide sequence revealed that the putative open reading frame was 511 base pairs and contained 169 amino acid residues. It is 79% and 57% identical to the rice cDNA (C26347) in DataBank at the nucleotide and amino acid sequence levels, respectively. The clone OPT8(511) specifically amplified a 511 bp band from the DNA of cold sensitive cultivars. Use of this marker could facilitate early selection of character associated with cold tolerance in rice. 相似文献
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We have developed a polymerase chain reaction (PCR)-based assay that could effectively reduce the time period required to screen and select the cold tolerance gene of rice seedlings under field conditions. The two specific random amplified polymorphic DNA (RAPD) fragments for the assay were identified on the basis of quantitative trait loci (QTL) analysis which were found to be tightly linked to cold sensitivity. The two RAPD fragments, OPT8(600) in the cold sensitivity rice cultivar 'Dular (indica)' and OPU20(1200) in the resistance rice cultivar 'Toyohatamochi (japonica)', were identified after screening 11 RAPD fragments using 2 random primers on the genomic DNAs of 'Dular' and 'Toyohatamochi'. These primers, when used in a multiplexed PCR, specifically amplified a 0.6 kb and a 1.2 kb fragment in the sensitive and resistant rice cultivars, respectively. When this assay was performed on the genomic DNAs of 16 japonica, 3 Tongil (indica/ japonica), and 2 indica rice cultivars, the primers amplified a 0.6 kb fragment in all of the cold sensitivity rice cultivars or 1.2 kb fragment in all of the resistance ones. These markers can be of potential use in the marker-assisted selection (MAS) for cold tolerance in rice seedling. As screening for resistance can now be conducted independent of the availability of low temperature, the breeding of cold tolerance cultivars can be hastened. 相似文献
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02428×合系35 RILs群体糙米和发芽糙米γ-氨基丁酸、抗性淀粉的遗传分析 总被引:1,自引:0,他引:1
以水稻02428×合系35的RILs群体及其亲本糙米和发芽糙米为材料,对糙米、发芽糙米各222个群体样品的γ-氨基丁酸、抗性淀粉含量进行测定,以期选育出高γ-氨基丁酸、高抗性淀粉的水稻新品种。研究结果表明,重组自交系糙米、发芽糙米γ-氨基丁酸含量差异不大,群体间存在广泛变异,由主效基因控制。重组自交系糙米、发芽糙米抗性淀粉含量差异大,发芽糙米抗性淀粉平均含量是该群体糙米的1.2倍,群体间存在广泛变异,呈偏态分布。高海拔冷凉气候有利于糙米高抗性淀粉含量的提升与进化。本研究可以为功能水稻的遗传及品种选育提供一定的理论依据。 相似文献
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A novel mutated acetolactate synthase gene conferring specific resistance to pyrimidinyl carboxy herbicides in rice 总被引:3,自引:0,他引:3
Acetolactate synthase (ALS) is the first common enzyme in the biosynthetic pathway of branched-chain amino acids. Mutations
of specific amino acids in ALS have been known to confer resistance to ALS-inhibiting herbicides such as sulfonylureas and
pyrimidinyl carboxy (PC) herbicides. However, mutations conferring exclusive resistance to PC have not yet been reported to
date. We selected PC resistant rice calli, which were derived from anther culture, using one of the PCs, bispyribac-sodium
(BS), as a selection agent. Two lines of BS-resistant plants carrying a novel mutation, the 95th Glycine to Alanine (G95A),
in ALS were obtained. In vitro ALS activity assay indicated that the recombinant protein of G95A-mutated ALS (ALS-G95A) conferred
highly specific resistance to PC herbicides. In order to determine if the ALS-G95A gene could be used as a selection marker
for rice transformation, the ALS-G95A gene was connected to ubiquitin promoter and introduced into rice. PC resistant plants
containing integrated ALS-G95A gene were obtained after selection with BS as a selection agent. In conclusion, novel G95A
mutated ALS gene confers highly specific resistant to PC-herbicides and can be used as a selection marker. 相似文献
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Jia L Yan W Zhu C Agrama HA Jackson A Yeater K Li X Huang B Hu B McClung A Wu D 《PloS one》2012,7(3):e32703
Sheath blight (ShB) caused by the soil-borne pathogen Rhizoctonia solani is one of the most devastating diseases in rice world-wide. Global attention has focused on examining individual mapping populations for quantitative trait loci (QTLs) for ShB resistance, but to date no study has taken advantage of association mapping to examine hundreds of lines for potentially novel QTLs. Our objective was to identify ShB QTLs via association mapping in rice using 217 sub-core entries from the USDA rice core collection, which were phenotyped with a micro-chamber screening method and genotyped with 155 genome-wide markers. Structure analysis divided the mapping panel into five groups, and model comparison revealed that PCA5 with genomic control was the best model for association mapping of ShB. Ten marker loci on seven chromosomes were significantly associated with response to the ShB pathogen. Among multiple alleles in each identified loci, the allele contributing the greatest effect to ShB resistance was named the putative resistant allele. Among 217 entries, entry GSOR 310389 contained the most putative resistant alleles, eight out of ten. The number of putative resistant alleles presented in an entry was highly and significantly correlated with the decrease of ShB rating (r = −0.535) or the increase of ShB resistance. Majority of the resistant entries that contained a large number of the putative resistant alleles belonged to indica, which is consistent with a general observation that most ShB resistant accessions are of indica origin. These findings demonstrate the potential to improve breeding efficiency by using marker-assisted selection to pyramid putative resistant alleles from various loci in a cultivar for enhanced ShB resistance in rice. 相似文献
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With the purpose of selecting and breeding cold stress tolerance of hybrid rice cultivars, the effect of chilling on five hybrid rice combinations was investigated. The results indicated that the cold tolerance of different hybrid rice varieties were different. The order of the cold tolerance as the degrees of inhibition of maximum photosynthetic rate and of apparent quantum yield of flag leaves at primary heading stage by chilling was as follows: The cold tolerance of Japanica type Xiuyou-57 was the highest then lndica type Qing-Youzao followed by Shangyou-63, Shangyou-64 and Weiyou-64. The cold tolerance of Fx hybrid rice was similar to that of the maternal lines and not similar to the paternal lines. The cold tolerance of maintainer lines was similar to that of the male sterile lines. The tests on the survival rate of hybrid rice seedlings after chilling treatment also showed a similar re gular patterns. These indicated that the responses by the chilling temperature in these five hybrid combinations were exactly the same in the Seedling stage as well as in the late growing stage. The results of these experiments on the impact of selecting and breeding cold tolerance in hybrid rice were discussed. 相似文献