抗生素对火炬松的器官发生植株再生的影响(英文)

本文研究了氨苄青霉素、羧苄青霉素和头孢霉素等3种抗生素对火炬松愈伤组织的生长和分化及不定芽生根的影响。结果表明,头孢霉素最有利于愈伤组织的诱导和生长,羧苄青霉素最有利于芽的分化,氨苄青霉素降低了不定芽的生根频率。所有试验的3种抗生素提高愈伤组织的形成和芽再生,但降低了芽的生根频率。这些结果表明,选择合适的抗生素对优化火炬松遗传转化体系有重要作用。图3表4参25。     

农杆菌介导的杜仲叶片愈伤组织遗传转化体系

【目的】优化杜仲叶片愈伤组织的再生体系,研究愈伤组织对抗生素和抑菌剂的敏感性,探索影响农杆菌介导杜仲愈伤组织遗传转化的最适转化因子水平,构建农杆菌介导的杜仲愈伤组织瞬时转化体系,使杜仲成年植株外植体为受体的遗传转化成为可能,为杜仲基因功能的研究与定向改良奠定基础。【方法】以杜仲成年植株叶片为材料诱导愈伤组织,通过添加不同浓度植物生长调节剂与大量元素的MS培养基进行不定芽的诱导与增殖,确定最适培养基。在此基础上,在培养基中添加不同浓度抗生素及抑菌剂,研究愈伤组织对其敏感性。以获得的叶片愈伤组织受体系统为基础,通过L_(16)(4~5)的正交试验,探索不同转化因子对农杆菌介导叶片愈伤组织遗传转化的瞬时转化效率的影响,建立最适瞬时转化体系。使用获得的瞬时转化体系对愈伤组织进行遗传转化操作,筛选抗性芽,对抗性芽进行GUS组织化学染色与PCR检验。【结果】再生体系优化的结果表明,3/4大量元素浓度的MS培养基能够促进杜仲愈伤组织不定芽的诱导及生长;愈伤组织诱导不定芽的最适培养基为3/4MS+0. 27μmol·L~(-1)NAA+4. 4μmol·L~(-1)6-BA,诱导率为83%±10. 0%;不定芽复壮的最适培养基为3/4MS+0. 054μmol·L~(-1)NAA+4. 4μmol·L~(-1)6-BA,平均伸长长度为(2. 47±1. 33) cm。抗生素与抑菌剂敏感性试验表明,遗传转化的选择培养基中,抑菌剂头孢霉素的最适浓度为200 mg·L~(-1),筛选用的抗生素卡那霉素最适浓度为70 mg·L~(-1)。转化因子的正交试验表明,最适的农杆菌介导杜仲叶片愈伤组织遗传转化的转化因子组合为:预培养5天、侵染10 min和共培养3天。使用最适瞬时转化体系对约200个愈伤组织进行遗传转化操作,共筛选获得3个抗卡那霉素的抗性芽; GUS组织化学染色显示GUS基因在抗性芽中得到了表达,PCR检测证明这些抗性芽中存在NPTⅡ基因。【结论】杜仲叶片愈伤组织诱导不定芽的最适培养基为3/4MS+0. 27μmol·L~(-1)NAA+4. 4μmol·L~(-1)6-BA,不定芽复壮的最适培养基为3/4MS+0. 054μmol·L~(-1)NAA+4. 4μmol·L~(-1)6-BA。农杆菌介导的杜仲叶片愈伤组织瞬时转化体系为:预培养5天、侵染10 min和共培养3天,筛选培养基为3/4MS+0. 054μmol·L~(-1)NAA+4. 4μmol·L~(-1)6-BA+200 mg·L~(-1)Cef+70 mg·L~(-1)Km。利用此体系共获得3个抗性芽,PCR分析和GUS组织化学染色都表明T-DNA已整合到抗性芽基因组中。     

抗生素对山茶愈伤组织诱导和生长的影响

研究了卡那霉素、潮霉素对浙江红山茶、大花红花油茶愈伤组织的生长及对攸县油茶幼苗叶片诱导愈伤的影响,确定作为选择压力的卡那霉素质量浓度为50～60mg·L-1,潮霉素为25～30mg·L-1。在3种抑菌性抗生素中,羧苄青霉素促进愈伤组织诱导,对愈伤组织的生长没有影响,头孢霉素则有抑制作用,而羟氨苄青霉素轻微抑制愈伤组织的生长,但对愈伤组织的诱导影响不大。考虑到抑菌效果,适于在转基因研究中使用的羧苄青霉素的抑菌质量浓度为250mg·L-1,羟氨苄青霉素为125mg·L-1。     

四倍体刺槐、国槐和红叶石楠对根癌农杆菌菌株及不同抗生素敏感性研究

以含有RD29A目的基因和NPTⅡ筛选基因的根癌农杆菌菌株LBA4404、GV3101和EHA105侵染饲用型四倍体刺槐(Robinia pseudoacacia)、国槐(Sophora japonica L.)和红叶石楠(Davidson Photinia)的组培苗叶片,研究3个树种对根癌农杆菌菌株和不同抗生素(替门汀和头孢霉素)的敏感性、以及根癌农杆菌菌株对3个树种的侵染能力。结果表明,不同树种、不同根癌农杆菌株和不同抗生素种类均对转化效果产生明显影响,红叶石楠是最佳受体材料,3个菌株侵染后的平均抗性愈伤率为37.0%,其次为国槐,四倍体刺槐最低;根癌农杆菌菌株LBA4404侵染能力最强,3个树种的平均抗性愈伤率达到33.3%、平均分化率达到17.6%;替门汀对农杆菌侵染后的红叶石楠组培叶片抑菌效果最好。     

种坯处理、不同激素浓度在暗培养条件下对多年生黑麦草愈伤组织诱导的影响

以多年生黑麦草种子为外随体，研究了在暗培养条件下，不同浓度2，4—D、BA及种胚处理对愈伤组织诱导的影响，试验表明：(1)切胚处理对多年生黑麦草愈伤组织诱导起到很大的促进作用；(2)种胚愈伤组织的诱导宜采用MS 2，4—D 5mg/L的培养基；(3)种子愈伤组织的诱导与种子发芽率无关。     

国槐遗传转化体系的优化

采用根癌农杆菌介导的国槐叶盘转化法,在建立修饰的豇豆胰蛋白酶抑制剂基因(sck)转化体系过程中,对影响农杆菌转化频率的各种因素进行研究.结果表明:国槐叶片需在黑暗条件下在MS培养基上预培养5天,农杆菌菌株选用GV3101,农杆菌共培养3天较合适;农杆菌菌液浓度OD600约为0.7,感染时间10 min;侵染前的菌液和共培养基中添加200/μmol·L-1乙酰丁香酮(AS)对国槐遗传转化有明显的促进作用;抑制农杆菌生长的抗生素浓度以头孢霉素(Cef)500mg·L-1最好.PCR和Southern blotting检测证实sck基因已整合到国槐基因组中.     

芍药愈伤组织诱导及遗传转化技术的初步研究

利用正交设计法诱导形成愈伤组织,并探讨6-BA,NAA,2,4-D等外源激素对芍药成熟胚愈伤诱导的影响。通过根癌农杆菌介导法将衰老抑制基因(IPT)转入芍药的愈伤组织:共处理72个愈伤组织块,用含除草剂的D4培养基进行选择培养,筛选出18个生长和增殖良好的愈伤组织块;经PCR鉴定,确认其中17个愈伤组织块中导入进IPT基因。     

农杆菌介导的楸树遗传转化体系

【目的】以楸树胚性愈伤组织为受体,建立有效的楸树遗传转化体系,为今后楸树性状的遗传改良奠定基础。【方法】通过农杆菌EHA105介导以胚性愈伤组织作为外植体进行遗传转化,通过正交试验获得最优的遗传转化条件,进而将外源基因转入到楸树基因组中。【结果】在1/2 MS培养基中添加不同梯度浓度的卡那霉素(Kana)进行选择压力筛选,在添加了60 mg·L~(-1)Kana的1/2 MS培养基中,楸树胚性愈伤组织的分化率为0.00%,存活率仅为5.71%,因此确定60 mg·L~(-1)为遗传转化的选择压。采用正交设计L18(37)进行农杆菌介导的楸树遗传转化试验,通过GUS化学组织染色统计瞬时表达率,正交试验直观分析和单因素方差分析结果表明:在预培养时间为2天,采用农杆菌菌株EHA105、菌液浓度OD600值为0.7、添加乙酰丁香酮(AS)浓度为300μmol·L~(-1)、侵染时间为10 min,共培养时间为5天的条件下,农杆菌介导的转化效率最高,且对转化效率影响最大的2个因素是乙酰丁香酮浓度和预培养时间。对浸染后的胚性愈伤组织进行8个月的筛选培养,共获得32个抗性组织团,对其中15个增殖较多的抗性愈伤组织进行PCR检测,表明86.67%的抗性组织团中有外源基因整合到楸树基因组中。内源激素水平会对植物体细胞胚分化产生影响,细胞分裂素(CTK)和脱落酸(ABA)促进体胚发生,生长素(IAA)和赤霉素(GA)对体胚发生有抑制作用。通过测定内源激素可知,转基因的抗性组织中内源CTK和ABA水平显著低于野生型的楸树胚性愈伤组织,而内源IAA和GA则显著高于野生型胚性愈伤组织,推测内源激素水平可能是转基因抗性组织体胚分化能力比较差的原因。【结论】建立了农杆菌介导的楸树胚性愈伤组织的遗传转化体系,对筛选获得的15个抗性愈伤组织进行PCR检测,其中13个抗性愈伤组织中有外源基因的整合。内源激素水平的变化可能是导致楸树转基因抗性愈伤组织难以分化的原因。     

不同的抗生素对农杆菌EHA105（codA）和农杆菌EHA105（betA）的抑制作用

研究了国产头孢曲松钠（Ceftriaxone sodium）,头孢唑林钠（Cefazolin sodium）,头孢霉素（Cefotaxime sodium）类抗生素在不同浓度下（400mg/L,500mg/L,700 mg/L,900 mg/L,1000 mg/L）对农杆菌EHA105（codA）和农杆菌ZHA105（betA）的抑制效果;再次研究了将不同产地的药品以1：1的比例混合后对农杆菌EHA105（codA）和农杆菌ZHA105（betA）的抑制效果。最后将混合头孢中加入不同浓度的AgNO3溶液（其浓度分为1mg/L,1.5mg/L,2mg/L,2.5mg/L和3.0mg/L,共五个梯度）,测定其对农杆菌EHA105（betA）的抑制效果。将AgNO3引入本实验是鉴于文献报道说Ag＋有良好的杀菌效果,对大肠杆菌,黄金色葡萄球菌,白念珠菌杀灭率在99.9%以上,不同浓度的Ag＋杀菌效果也各不相同,因此在本实验中引入AgNO3,测定其对农杆菌的抑制效果如何。本次所做的抑菌实验采用了农杆菌EHA105（betA）和农杆菌EHA105（codA）,选择出了较好的菌种EHA105（betA）,在未加入AgNO3的抗生素中以哈曲混合药在500mg/L时的抑菌效果为最好,单一抗生素则以鼎国头孢霉素在500mg/L时的抑菌效果最好。在加入AgNO3的抗生素中,选择出了AgNO3的最佳抑菌浓度为2.5mg/L,抗生素为哈药的头孢曲松钠在加入1.0mg/L的AgNO3时有良好的抑菌作用。由于将药品混合后其抑菌效果并没有太大的变化和差异,所以建议不采用混合药品。     

樟树叶及天竺桂叶的精油抑菌活性研究

采用水蒸馏法提取樟树叶、落叶、花及天竺桂叶的精油,用杯碟法测定了2种植物叶精油对大肠杆菌、金黄色葡萄球菌、青霉和黑曲霉的抑菌活性,并测定了2种精油对应试菌种的最小抑菌浓度(MIC).结果表明;樟树的落叶含精油最高,可达0.52%,天竺桂鲜叶含精油量达0.53%;樟树叶精油对4种供试菌都有较好的抑菌效果,但天竺桂精油对青霉无抑制作用;樟树叶精油对大肠杆菌、金黄色葡萄球菌、青霉和黑曲霉的最小抑菌浓度(MIC)分别为2.500 μL/mL,2.500 μL/mL,0.625 μL/mL,1.250 μL/mL;天竺桂叶精油对大肠杆菌、金黄色葡萄球菌和黑曲霉的最小抑菌浓度(MIC)分别为1.25 μLm/L,2.50 μL/mL,5.00 μL/mL.     

红松不同部位精油的成分分析及抑菌活性

[目的]植物精油是安全并有良好抑菌效果的天然产物,在食品防腐剂领域潜力巨大.本研究通过对红松不同部位精油成分与抑菌活性的研究,能为开发安全绿色的食品防腐剂及为其在食品、化妆品等领域的应用提供理论基础.[方法]采用水蒸气蒸馏法从红松的松针、松塔、松壳中制备精油,采用气相色谱-质谱(GC-MS)联用技术结合保留指数(RI)...     

无瓣海桑对乡土红树植物的化感作用

以外来种无瓣海桑为供体,通过室内栽培实验探讨了无瓣海桑各器官水浸液的不同浓度(即0 1、0 2、0 3、0 4、0 5g·mL-1)对我国4个乡土红树植物的化感作用。结果表明:无瓣海桑对各乡土红树植物均存在化感作用,对胚轴萌发的影响大于对幼苗生长的影响;除最低浓度外,不同浓度的各器官水浸液均能明显抑制各乡土红树植物的萌发,且随着浓度增大抑制作用增强;所有水浸液均表现出高浓度下抑制幼苗生长,随着浓度的降低抑制作用减弱,甚至转为促进的规律;各器官的水浸液对各乡土红树植物的萌发和幼苗生长的抑制作用为:果>根>凋落叶;高浓度时,无瓣海桑的各器官水浸液对不同受体红树植物的幼苗生长抑制强度不同。     

Screening antiacne potency of Indonesian medicinal plants: antibacterial,lipase inhibition,and antioxidant activities

Indonesian medicinal plants were screened as potential sources of antiacne agents. The screening methods were performed using antibacterial assay against Propionibacterium acnes, lipase inhibitor assay, and antioxidant assay. The results showed that from 40 plant materials extracted with methanol and 50% ethanol in water, Caesalpinia sappan was the best extract based on the combined activities: antibacterial (minimum inhibitory concentration 0.13 mg/ml; minimum bactericidal concentration 0.25 mg/ml), lipase inhibitory [50% inhibitory concentration (IC₅₀) 120.0 μg/ml], and antioxidative (IC₅₀ 6.47 μg/ml). Another prospective extract is Intsia palembanica based on its lipase inhibitory activity (IC₅₀ 4.1 μg/ml) and antioxidant activity (IC₅₀ 3.87 μg/ml). Part of this report was presented at the 58th Annual Meeting of the Japan Wood Research Society, Tsukuba, March 2008     

Growth and physiological responses of chestnut calli to crude extracts of virulent and hypovirulent strains of Cryphonectria parasitica

The effect of crude extracts (CEs) of virulent (V‐E4) and hypovirulent (H‐E13) Cryphonectria parasitica strains on growth and physiological activities of chestnut calli was investigated on cell cultures obtained from either a susceptible Castanea sativa or a resistant Castanea mollissima chestnut species. The V‐E4 CEs significantly reduced calli fresh weight in both species when used at 50 and 500 μg g^–1 and, in general, showed an inhibitory effect on either O₂ uptake and H⁺‐ATPase activity. Only the highest (500 μg g^–1) concentration of H‐E13 CEs had a detrimental effect on callus growth, whereas the lowest one (5 μg g^–1) induced a slight, but significant early increase in callus fresh weight of both genotypes. The O₂ uptake and H⁺‐ATPase activity were enhanced by the presence of H‐E13 CEs. The decrease of callus growth and physiological activities, caused by V‐E4 CEs, was attributed to toxic compounds produced by the C. parasitica virulent strain, whereas the stimulatory effects of H‐E13 CEs on the overall callus metabolism seemed to be due either to the lack or very reduced amount of phytotoxic compounds and to growth‐regulating substances produced only by the hypovirulent strain of the fungus.     

Influences of antibiotics on plantlet regeneration via organogenesis in Ioblolly pine （Pinus taeda L.）

Three antibiotics ampicillin, carbenicillin, and cefotaxime were evaluated for their effects on induction, growth, and differentiation of organogenic calli, as well as rooting of regenerated shoots of three Ioblolly pine (Pinus taeda L.) genotypes. Of the antibiotics administered, cefotaxime maximally increased the frequency of callus formation and growth rate of organogeni ccalli, carbenicillin maximally increased the frequency of shoot regeneration and the average number of adventitious shoots per piece of organogenic callus, ampicUlin maximally decreased the rooting frequency of regenerated shoots and mean number of roots per regenerated shoot, in comparison with antibiotic-free media. Compared with the control, ampicillin minimally increased the frequency of callus formation, cefotaxime minimally increased the frequency of shoot regeneration, and carbenicillin minimally decreased the rooting frequency of regenerated shoots in three Ioblolly pine genotypes tested. All three antibiotics increased the frequencies of callus formation and shoot regeneration, and reduced the rooting frequency ot regenerated shoots suggested that the establishment of an efficient Agrobacterium tumefaciens-mediated transformation protocol for stable integration of foreign genes into Ioblolly pine need to select a suitable antibiotic. This investigation could be useful for optimizing genetic transformation of conifers.     

含羞草叶子提取物对常见农作物种子萌发生长的抑制作用

观察了含羞草叶子提取物对孟加拉常见农作物种子萌发生长的抑制作用.实验在27-30℃的室温和24小时的光照条件下进行.结果表明：不同浓度的含羞草叶子提取物明显影响所选的植物种子根,茎和侧根的延长和生长.生物学测定表明：高浓度（50%-100%）的提取物有较强的抑制作用.而低浓度 （10%-25%） 的提取物的抑制作用较弱.研究还显示,提取物对植物根和侧根的抑制作用比对种子萌发和茎的生长抑制作用更显著.     

Antibacterial activity of Nepalese wild mushrooms against <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> and <Emphasis Type="Italic">Propionibacterium acnes</Emphasis>

The present study aims to reveal the antibacterial potential of the wild mushrooms of Nepal. Despite the recognition of the medicinal potential of the natural resources in this country, a systematic study on the bioactivities of the wild mushrooms is still lacking. Therefore, in an attempt to fill this gap, ethanol extracts of 90 Nepalese wild mushroom samples were tested for antibacterial activity against Staphylococcus aureus and Propionibacterium acnes. Staphylococcus aureus was comparatively more susceptible with Inonotus clemensiae exhibiting the least minimum bactericidal concentration (MBC) of 100 μg/mL. The major compound in I. clemensiae was identified to be hispidin using high resolution liquid chromatography–electron spray ionization mass spectrometry (LC–ESI-MS) and nuclear magnetic resonance (NMR) analyses. Furthermore, the minimum inhibitory concentration (MIC) and MBC values of hispidin were determined to be 25 and 100 µg/mL for S. aureus and P. acnes, respectively. These findings show that the Nepalese wild mushrooms have the potential to be a novel addition to the functional ingredients industry due to their strong antibacterial potential.     

猕猴桃采后病害植物源杀菌剂的筛选及其抑菌效果分析

【目的】为寻找对猕猴桃果实软腐病主要病原菌具有较佳抑菌效果的植物源保鲜剂(植物提取物),以促进猕猴桃产业健康、可持续的发展。【方法】以猕猴桃葡萄座腔菌和间座壳菌为供试真菌,选择120种具有抗菌抑菌作用的药用植物的丙酮提取物,采用牛津杯法对其离体抑菌活性进行了测定。【结果】在供试药材(生药)提取物的质量浓度为1.0 g/mL的条件下进行初筛,得到了20余种具有较强抑菌作用的植物品种;在以丁香、肉桂、零陵香、黄芩、油茶粕、广藿香、石菖蒲、迷迭香、黄连、凤仙透骨草、山苍子等植物的提取物处理后,2种病原真菌的抑菌圈直径均可达15 mm以上;其中丁香和肉桂的抑菌效果最好,其生药提取物的最低抑菌浓度和最低杀菌浓度均较低,分别为3.125与62.5 mg/mL;其次是细辛、葡萄籽、川芎、毛蕨、苦参、麻黄根、皂角刺、荆芥、独活、杜仲、五加皮等物种,其生药提取物的最低抑菌浓度和最低杀菌浓度分别约为10.0、150 mg/mL。当肉桂、丁香、黄芩提取物的浓度达到2.0 mg/mL时,此3种植物提取物处理组的间座壳菌菌丝干质量均在0.05 mg以下,而其葡萄座腔菌菌丝干质量均在0.1 mg以下;当提取物的处理浓度达到2.0 mg/mL时,肉桂、丁香、黄芩提取物处理组的果实发病率与对照之间分别大约相差31.94%、41.24%、37.60%,说明各处理均能有效降低猕猴桃果实的发病率。【结论】植物提取物作为一种新型植物源保鲜剂为猕猴桃果实长期高效、绿色、环保贮藏提供了方向,具有一定的实用意义。     

Effects of plant growth regulators,carbon sources and pH values on callus induction in Aquilaria malaccensis leaf explants and characteristics of the resultant calli

The endangered tropical tree, Aquilaria malaccensis, produces agarwood for use in fragrance and medicines. Efforts are currently underway to produce valuable agarwood compoundsn tissue culture. The purpose of this study was to develop an optimal growth medium, specifically, the best hormone combination for callus suspension culture. Using nursery-grown A. malaccensis, sterilized leaf explants were first incubated on basic Murashige and Skoog(MS) gel medium containing 15g/L sucrose and at pH 5.7. Different auxin types including 1-naphthaleneacetic acid(NAA), 2,4-dichlorophenoxyacetic acid(2,4-D), and indole-3-butyric acid(IBA), were tested at various concentrations(0.55, 1.1 and 1.65 μM) using the basic medium. Leaf explants were incubated for 30 days in the dark. Callus induced by 1.1 μM NAA had the highest biomass dry weight(DW) of 17.3 mg; however the callus was of a compact type. This auxin concentration was then combined with either 6-benzylaminopurine(BAP) or kinetin at 0.55, 1.1, 2.2 or 3.3 μM to induce growth of friable callus. The 1.1μM NAA + 2.2μM BAP combination produced friable callus with the highest biomass(93.3mg DW). When testing the different carbon sources and pHs, sucrose at 15g/L and pH at 5.7 yielded highest biomasses at 87.7mg and 83 mg DW, respectively. Microscopic observations revealed the arrangement of the friable cells as loosely packed with relatively large cells, while for the compact callus, the cells were small and densely packed. We concluded that MS medium containing 15 g/L sucrose, 1.1 μM NAA + 2.2 μM BAP hormone combination, and a pH of 5.7 was highly effective for inducing friable callus from leaf explants of A. malaccensis for the purpose of establishing cell suspension culture.