309 resultados para PL


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利用发根农杆菌(Agrobacterium rhizogenes)1601,1000,1500,15834,A4,均成功地转化了中药青蒿(Artemisia annua L.)并且建立了pRi1601,pRi15834,pRiA4诱导的发根培养。pRi1601,pRi15834的发根诱导率比其它质粒高。太老或太幼的叶片不利子发根的诱导;发根主要从叶脉的伤口处萌发;带顶芽或带侧芽的叶片容易诱导根,但不一定是发根。光照有利于发根的诱导和发根的生长。以每个发根的“绝对生长速率”(Gtowth Ratio,GR)和绝对“侧根”数量(Number of Side Roots,NSR),通过大量的发根系的筛选,建立了8个发根系,1601-L-1, 1601-L-2, 1601-L-3, 1601-L-4, 15834-L-1, 1601-P-I, 16 01-P-2,15834-L-2。Southern分子检测表明,160l-1-1,1801-L-2, 1601-L-3,1601-L-4,1601-P-1,1601-P-2均为转化子。8个建立的发根系之间无论生长或者QHS的合成存在明显的差异。比较光/暗(16/8hrs),25℃条件下培养的16 01-L-1,1601-L-2,1601-L-3,1601-L-4,1601-P-l,和1601-P-2,其中16 01-L-3的生长最快,160l-L-1的生长最慢;但是,1601-L-1的QHS的含量最高(可达1. 048%),1601-1-3的QHS的含量最低。160Z-L-3,15834 -L-1和2583:1-L-2的生长速率相差不大。用盛有l000mLMS液体培养基的3000mL的锥形瓶扩大培养1601-L -3,15834-L-1和15834-L-2,转速为ll0rlpm,培养过程中发根容易形成发根球(Hairy Root Balis,HRB),HRB的形成严重影响发根的生长和QHs的合成,HpLC分析表明扩大培养发根中QHS的含量比较低。 改变MS基本培养基中的无机离子的浓度,研究不同无机离子对发根生长和QHS的合成的影响。 l、KN03为18.79×10-3M时有利于1601- L-1生长,为14. 84×10-3M时有利于QHS的合成。NH-4N0-3浓度在10.93-12. 49×10—3M范围内有利于1601-L-1生长,在0-20.62×10-3M范围内对QHS的合成影响不大,大于20. 62×lO-3M不利QHS的合成。培养基中NH-4+/N0-3-比值为0. 37-0. 4-0.52:1时有利于发根的生长,比值为0.52 - 0.58:1时有利于QHS的合成。 2、H-2P0-4-浓度为2.498×10-3M时有利于发根的生长在0-2. 498×l0-3M范围内,随着浓度的提高,促进发根的生长。培养基中的H2P4 -的浓度在0-1.249×lO-3M的范围内,随着浓度的提高,促进QHS的合成,为1.249×10-3M时QHS的含量最高。 3、培养基中最适16 01-L-1生长的Ca-2+浓度为0.198- 0.766×10-3M,大于或小于该浓度范围,显著地抑制发根的生长。但是,在0-3.695×10-3M范围内,随着培养基中Ca-2+浓度提高,促进QHS的合成,最适Ca-2+浓度为3.695×l0-3M。 4、培养基中不加Mg-2+时,完全抑制发根生长,在0. 142×10-3M-7.506×l0-3M浓度范围内,对发根生长影响没有明显的差别。但是,HPLC和UV分析发根中QHS含量,培养基中不加Mg-2+时,发根中QHS含量最高。 5、培养基中的Fe-2+浓度在0. 25 -1.0×10-3M范围内,同时有利于16 01- L-1的生长和QHS的形成。 6、培养基中最适合予16 01- L-3生长的KI浓度为2.5ppm,大于或小予该浓度均显著地抑制发根的生长,培养基中加入KI明显地降低发根中的QHS的含量。 7、H2BO3对l601-L-l生长影响不大,HPLC分析QHS的含量,培养基中的H3BO3浓度为100ppm和400ppm,QHS的含量分别为1.69mg/g和1.80mg/g(DW)。 8、Cu-2+对1601-L-3的生长影响显著,最适合1601-L-3生长的Cu-2+浓度为1.00ppm,在0 -1.00ppm的浓度范围内,随着培养基中的Cu+浓度的提高,发根的生物量不断增加。培养基中QHS合成的最适Cu2+浓度为0.05ppm,大于或小于该浓度均显著地抑制发根中QHS的合成。 比较光培养和暗培养对发根生长的影响,结果表明光照明显地促进1601-L-l的生长,暗培养明显不利于发根的生长。最适合于发根生长的温度为25℃,大于35℃显著地抑制发根的生长,影响发根的根尖细胞的正常分裂。 改变培养基中的蔗糖浓度和在发根培养的不同时期给培养基中添加蔗糖,试验结果表明蔗糖作为碳源对1601-L-3和1601-L-1的生长具有显著的影响。 (1)培养基中缺少蔗糖显著地抑制发根的生长。 (2)发根培养的前5天时间内,蔗糖浓度为30- 60glL昀培养基最有利于发根的生长,50glL的培养基中的发根生长最快,培养基中的蔗糖浓度大于60g/L小于30g/L时,发根的生物量增加较少。 (3)发根培养至第15天时,蔗糖浓度为60g/L的培养基最有利予发根的生物量的增加。发根培养至30天时,蔗糖浓度为60-90g/L的培养基,发根的生物量的增加相差不大,但是为蔗糖浓度为30-40g/L的培养基中的发根生物量一倍。 (4)发根培养过程中,分别于第5和15天给蔗糖浓度为30g/L的培养基中添加一次或二次蔗糖,使培养基中的蔗糖终浓度相当于60g/L或90g/L,培养至30天时,添加蔗糖的培养基中的发根的干重生物量相当于不添加蔗糖培养基中的发根生物量一倍,相当于初始蔗糖浓度为60g/L和90g/L培养基中发根的生物量。 (5)随着培养基中蔗糖浓度的提高,发根干重/鲜重比显著增加。培养基中的蔗糖的消耗量与发根生物量的增加呈正相关,蔗糖消耗越多,发根生物量的增加越大。 比较pH值对发根生长和QHS合成的影响表明,灭菌前pH值在5.O-6.5范围内的培养基适合予1601-L-1的生长,小于5.O不利于发根的生长,pH5.8有利于1601-1-1生长和QHS的生物合成。发根收获时培养基中的pH值一般为4.5-5.2. pH7.O抑制发根的生长,pHl0.O对发根具有强烈的致死作用。发根在培养过程中,对培养基中的pH值具有显著的调节作用,发根能在很短的时间内(24- 48hrs)使pl:l值为5.8、6.4、7.0培养基降低到pH4. 5-5.2,pH为5.8的培养基有利于QHS合成。 比较不同基本培养基对发根生长和QHS合成的影响,试验结果表明N6、DCR、Litvay培养基有利于1601-L-1的生长,WS、White、B5培养基不利于发根的生长。DCR培养基中的QHS含量最高。 根据三水平试验选用三水平正交表来安排试验的原则,选用三水平正交表L7(3-),研究多因子效应对发根生长和QHS合成的影响,试验结果表明,Mg2+,Fe2+,Mn-2+,NH4NO3,KN03 ,KI,Ca-2+为发根生长的主要因子,NH4N03,KNOs,Mg2+,Ca2+,肌醇为QHS合成的主要因子。 通过TLC分析发根中QHS和其它化学成分,同时比较发根和无菌苗及野生植株的化学成分,发根和无菌苗均能合成包括QHS在内的野生青蒿叶片中的大部分非挥发性的化台 物。 研究青蒿植株在发育过程中QHS的含量的变化以及发根、无菌苗和野生青蒿中QHS的合成,HP分析结果表明,l、不同的单株青蒿之间的QHS量相差很大。2、同一植株幼 叶的QHS含量比老叶的QHS含量高。3、不同单株青蒿之间达到最高QHS含量的时间不一样,开花期或开花之前。4、无菌苗(带根)或者不带根丛生芽均能合成QHS,但是带根的无菌蕾的QHS量比丛生芽中的QIS的含量高。5、不同发根农杆菌转化的发根系1601-L-1和15834-L-1都能合成QHS。

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近年来植物重金属的耐性机制研究及抗重金属基因工程取得了很大进展。本文将来自于菜豆(Phaseolus vulgaris)的特异性重金属胁迫相关基因PvSR2 (Phaseolus vulgaris stress-related protein, PvSR)的cDNA序列克隆到大肠杆菌高效表达载体pBV221的PR PL启动子的下游,构建了原核表达载体pBV221-PvSR2。通过温度诱导,在大肠杆菌中成功地高效表达了PvSR2基因。经重金属(CdCl2)抗性检测,实验组比对照组有明显的抗性。 同时,将该基因克隆到植物转达化中间载体pCAMBEIA2301的花椰菜花椰病毒的35S启动子下游,利用根癌农杆菌(Agrobacterium tumefaciens)Ti质粒介导的遗传转化系统,成功地将该基因导入了烟草的基因组,获得了转基因植株。

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凋落物的分解对生物地球化学循环起着重要的作用。本研究以位于中国北方农牧交错区(内蒙古多伦县)的半干旱克氏针茅(Stipa krylovii Roshev.)草原生态系统为背景,针对凋落物分解研究中的几个关键过程和问题进行了探讨。研究内容包括该生态系统中几种常见植物凋落物的分解速率,在分解过程中的养分动态,凋落物的混合效应(即非加性效应)及其机制,全球变化引起的土壤有效N、P和降水增加对凋落物分解的影响及其对草原生态系统碳储量的影响等。 通过对克氏针茅、糙隐子草(Cleistogenes squarrosa Trin.)、双齿葱(Allium bidentatum Fisch.)、野韭(Allium ramosum L. Sp. Pl.)和冰草(Agropyron cristatum Gaertn.)的叶、茎、根凋落物的研究发现,叶和茎的分解速率与凋落物初始N和P含量呈显著正相关,根系的分解速率与初始C/N比呈显著负相关。根系凋落物一般比叶和茎凋落物分解速率快,而且具有更快的养分周转速率。有些凋落物混合在一起分解后对分解速率产生了混合效应(混合凋落物的实际分解速率偏离于基于组分凋落物计算的预期分解速率),同时它们对养分(主要是N、P、Ca、Mg)的固定或释放被延后。混合效应发生与否以及混合效应的方向主要取决于组分凋落物的特点,而与凋落物多样性的高低没有明显的关系。混合凋落物对养分动态的负作用,能降低退化生态系统遭到干扰后养分的损失,而且养分释放时间的延后有助于某些植物更好地生长,有利于退化草原生态系统恢复过程中植被结构的改善。 由于凋落物的质量对分解速率的影响重大,凋落物的化学组成很可能对混合凋落物分解过程中的非加性效应起重要作用,然而迄今为止没有明确的结论。本文研究发现,具有不同初始N、P含量的凋落物(同一物种)混合在一起分解后产生了正的非加性效应,非加性效应的强弱与组分凋落物初始N、P含量的差异大小有关。而且,非加性效应与P含量差异大小的相关性比与N的强,这可能与研究地点土壤中P的含量相对更为缺乏有关。这一研究结果表明,凋落物的化学组成在有些情况下确实与非加性效应有关。 通过施肥和浇水试验,研究了全球变化引起的土壤有效N、P和水分的增加对具有不同生活型和化学组成的两种优势草原植物,即双齿葱和克氏针茅凋落物分解的直接影响。结果发现,向土壤中添加N肥或N、P复合肥,在短期内(100天)加速了这两种植物的分解速率。高质量凋落物(双齿葱)的分解更容易受到土壤水分条件的限制,而低质量凋落物(克氏针茅)的分解对土壤养分有效性更敏感。土壤中有效养分的增加会提高分解凋落物对养分的固定,有利于退化生态系统中养分的保持。双齿葱对内蒙古半干旱典型草原的碳循环和养分动态的贡献,比克氏针茅要更大。研究结果提示人们,对草原生态系统的碳循环和养分动态进行模拟时,需要对不同化学组成的凋落物分别加以考虑。 本文还探讨了土壤有效N长期增加后对两种优势草原植物(克氏针茅和冷蒿(Artemisia frigida Willd))凋落物分解的综合影响,并进一步区分了由土壤有效N增加引起的凋落物质量改变的间接作用和土壤状况改变的直接作用。结果发现,土壤有效N长期增加后对凋落物的分解速率没有明显影响,但是显著加速了分解凋落物对养分的固定,这可能是由于凋落物质量对分解的促进作用和土壤状况对分解的抑制作用相互抵消的原因所造成的。向土壤中添加N素后,地上部植被生产力不可避免的会提高,尽管短期内会加速凋落物的分解速率,但从长远来看,对凋落物的分解速率不会产生影响,所以估计土壤有效N增加后最终会提高草原生态系统的净碳储量。 通过本研究,对中国北方农牧交错区(内蒙古多伦县)半干旱克氏针茅草原生态系统的养分循环过程有了初步的了解,验证了混合凋落物分解过程中非加性效应发生的可能性,并且证明组分凋落物的初始化学组成对这种非加性效应确实起着重要作用。本研究说明全球变化对不同植物种凋落物分解的影响是不同的,而且因研究时间长短的不同对凋落物分解的影响会发生变化。研究结果为草原生态系统碳和养分循环过程的模拟提供了必要的基础资料,为混合凋落物分解过程中非加性效应发生的可能机制提供了强有力的证据,对在全球变化背景下人们对草原生态系统碳和养分循环过程的认识和模拟有着重要的参考价值。

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光敏核不育水稻农垦58S是石明松于1973年在晚粳农垦58的大田中发现的雄性不育突变体,它在长日照下雄性不育可被用于与恢复系杂交生产杂种,而在短日照下雄性可育能用于自交繁殖,它的恢复系来源广泛。基于这些特性,育种学家用光敏核不育水稻建立的二系杂交水稻制种技术有很大的应用潜力。近十几年来,育种学家用农垦58S作基因供体转育了许多新的不育系,研究结果表明育成的粳型不育系均为光敏不育系,但在育成的籼型不育系中,绝大多数丧失光敏核不育特性,变成温敏不育系。目前因不知光敏核不育的分子遗传机制,尚不能解释这些问题。 本文用双向电泳技术分析了农垦58S和农垦58苗期和育性转换光敏感期叶绿体蛋白质的差异,在农垦58S中发现三个蛋白质(Pl,P2和P3),其中Pl和P2在苗期和光敏感期叶片内均存在,P3仅在光敏感期的叶片中存在,它们不受长日照或短日照处理的影响。农垦58没有这三个蛋白质。 用制备型双向电泳纯化后,得到SDS - PAGE和IEF纯的Pl和P2。经SDS-PAGE和IEF测定,Pl的等电点是6.2,分子量是41 kDa;P2的等电点是5.8,分子量是61 kDa。现称Pl为P41,P2为P61。氨基酸序列分析和同源性检索发现P41与水稻叶绿体ATP合成酶p亚基和酵母转录因子CAD1有同源性,此外,P41的N-端序列中有一个与蛋白激酶催化核心中的多功能motif Y-G-X-G-X- (P/T)-G-V相似的序列;P61的14个氨基酸长的N-端序列与水稻叶绿体ATP合成酶β亚基的一致。P41和P61 N-端前12个氨基酸的序列也完全一致。 PCR扩增和Southern杂交分析没有发现农垦58S和农垦58之间ATP合成酶β亚基基因(atpB)的多态性。Nothern杂交分析表明农垦58S中仅有一种、与农垦58 atpB mRNA分子量相同的atpB转录产物,但它的atpB mRNA丰度明显低于农垦58的。没有检测到突变的atpB和其它形式的atpB转录产物。 分析P41和P61在其它水稻材料中的分布特点发现它们在粳型光敏不育系7001S、5088S、31301S、C407S和1647S,籼型光敏不育系W7415S和W9451S以及温(光)敏不育系培矮64S中存在,而在对照材料三系水稻马协A、珍汕97A、马协B、珍汕97B和明恢63以及常规粳稻C94153中不存在。根据这些不育系的系谱和它们与农垦58S之间基因的等位性研究结果,讨论了P41和P61与光敏核不育性的可能联系。

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We have made a set of chromosome-specific painting probes for the American mink by degenerate oligonucleotide primed-PCR (DOP-PCR) amplification of flow-sorted chromosomes. The painting probes were used to delimit homologous chromosomal segments among human, red fox, dog, cat and eight species of the family Mustelidae, including the European mink, steppe and forest polecats, least weasel, mountain weasel, Japanese sable, striped polecat, and badger. Based on the results of chromosome painting and G-banding, comparative maps between these species have been established. The integrated map demonstrates a high level of karyotype conservation among mustelid species. Comparative analysis of the conserved chromosomal segments among mustelids and outgroup species revealed 18 putative ancestral autosomal segments that probably represent the ancestral chromosomes, or chromosome arms, in the karyotype of the most recent ancestor of the family Mustelidae. The proposed 2n = 38 ancestral Mustelidae karyotype appears to have been retained in some modern mustelids, e.g., Martes, Lutra, ktonyx, and Vormela. The derivation of the mustelid karyotypes from the putative ancestral state resulted from centric fusions, fissions, the addition of heterochromatic arms, and occasional pericentric inversions. Our results confirm many of the evolutionary conclusions suggested by other data and strengthen the topology of the carnivore phylogenetic tree through the inclusion of genome-wide chromosome rearrangements. Copyright (C) 2002 S. KargerAG, Basel.

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The Afrotheria, a supraordinal grouping of mammals whose radiation is rooted in Africa, is strongly supported by DNA sequence data but not by their disparate anatomical features. We have used flow-sorted human, aardvark, and African elephant chromosome painting probes and applied reciprocal painting schemes to representatives of two of the Afrotherian orders, the Tubulidentata (aardvark) and Proboscidea (elephants), in an attempt to shed additional light on the evolutionary affinities of this enigmatic group of mammals. Although we have not yet found any unique cytogenetic signatures that support the monophyly of the Afrotheria, embedded within the aardvark genome we find the strongest evidence yet of a mammalian ancestral karyotype comprising 2n = 44. This karyotype includes nine chromosomes that show complete conserved synteny to those of man, six that show conservation as single chromosome arms or blocks in the human karyotype but that occur on two different chromosomes in the ancestor, and seven neighbor-joining combinations (i.e., the synteny is maintained in the majority of species of the orders studied so far, but which corresponds to two chromosomes in humans). The comparative chromosome maps presented between human and these Afrotherian species provide further insight into mammalian genome organization and comparative genomic data for the Afrotheria, one of the four major evolutionary clades postulated for the Eutheria.

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Multidirectional comparative chromosome painting was used to investigate the karyotypic relationships among representative species from three Feliformia families of the order Carnivora ( Viverridae, Hyaenidae and Felidae). Complete sets of painting probes derived from flow-sorted chromosomes of the domestic dog, American mink, and human were hybridized onto metaphases of the spotted hyena (Crocuta crocuta, 2n = 40) and masked palm civet (Paguma larvata, 2n = 44). Extensive chromosomal conservation is evident in these two species when compared with the cat karyotype, and only a few events of chromosome fusion, fission and inversion differentiate the karyotypes of these Feliformia species. The comparative chromosome painting data have enabled the integration of the hyena and palm civet chromosomes into the previously established comparative map among the domestic cat, domestic dog, American mink and human and improved our understanding on the karyotype phylogeny of Feliformia species. Copyright (C) 2005 S. Karger AG, Basel.

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To better understand the evolution of genome organization of eutherian mammals, comparative maps based on chromosome painting have been constructed between human and representative species of three eutherian orders: Xenarthra, Pholidota, and Eulipotyphla,

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在大鼠基因组数据库中搜索得到两个泌乳刺激素基因家族的新成员.进一步分析显示该基因家族起源于啮齿目和其他哺乳动物分歧之后,而且大部分基因座位的重排在大、小鼠分歧之前已经完成.但PL-Ⅰ和PL-Ⅱ基因簇却是例外,它们在基因树上以物种特异的方式聚类.结合基因转换的检验、染色体上相对位置比较和基因重复时间估计的结果,认为啮齿目PL-Ⅰ和PL-Ⅱ基因是物种特异的,它们由一系列在大、小鼠分歧之后发生的基因重复事件形成.结果还揭示了在啮齿目泌乳刺激素基因家族进化过程中持续不断的发生了基因重复和基因分化事件.

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采用PCR 方法从4 个灵长目物种中获得了泌乳刺激素(Prolactin, PRL)基因的第2~5 外显子的序 列, 结合GenBank 中已有的序列, 计算了PRL 基因在灵长目中的氨基酸替换速率. 对异义替换速率和 同义替换速率进行比较的结果显示, 在灵长目进化的过程中未发现任何一个枝系有正选择作用的迹象. 此外, 用最大似然(Maximum-Likelihood, ML)法没有检验到任何一个受到正选择作用的位点; 在快速 进化时期内发生的所有32 个氨基酸替换中, 只有2 个位点包含在功能重要的40 个氨基酸之内, 提示所 发生的氨基酸替换主要集中在功能上并不是很重要的位点上. 此外, 在PRL 基因的快速进化过程中, PRL 的部分功能被胎盘泌乳刺激素(placental lactogen, PL)所替代. 基于以上原因, 推测发生在灵长目 PRL 基因的快速进化很可能是选择压力的放松, 而不是正选择作用的结果.

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The 9-bp deletion in the COII/tRNA(Lys) intergenic region (region V) of human mitochondrial DNA was screened in 1521 Chinese from 16 ethnic groups and 9 Hen geographic groups. The highest frequency was found in populations of Miao (32.4%) and Bouyei (30.8

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Penaeus monodon postlarvae were fed with different percentages (0%, 25%, 50%, 75% and 100%) of the herbal appetizer Zingiber officinalis enriched Artemia. After 30 days of culture (i.e. PL-1-30), a very positive result was found in Z. officinalis-enriched Artemia-fed postlarvae. The unenriched Artemia-fed postlarvae consumed 91.0 mg/animal/30 days of feed, whereas the Z. officinalis-enriched Artemia increased their consumption to 127.9 mg/animal/30 days. A similar pattern was noticed in feed absorbed (110.2 mg), dry weight growth (26.7 mg) and feed catabolized (83.2 mg) in Z. officinalis-enriched Artemia because of enzymatic activities. The conversion efficiency of unenriched postlarva was 17.19%, whereas in 100% Z. officinalis-enriched Artemia, the maximum conversion efficiency was 20.85%. The net production efficiency increased significantly (P < 0.05) to 22% from that of the unenriched Artemia-fed postlarvae. The administration of Z. officinalis in all levels produced significantly (P < 0.05) higher weight gain and specific growth rate. The utilization efficiency of feed increased proportionately to the percentages of Z. officinalis. Digestive enzyme activity (amylase, protease and lipase) increased significantly (P < 0.05) in the 50%, 75% and 100% enrichment. Among the different percentages of enrichment, the 100% Z. officinalis-enriched Artemia-fed postlarvae performed better in the overall status.

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A phytoplankton-lytic (PL) bacterium, Bacillus cereus, capable of lysing the bloom-forming cyanobacterium. Aphanizomenon flos-aquae was isolated from Lake Dianchi of Yunnan province, China. This bacterium showed lytic activities against a wide range of cyanobacteria/algae, including A. flos-aquae, Microcystis viridis, Microcystis wesenbergi, Microcystis aeruginosa, Chlorella ellipsoidea, Oscillatoria tenuis, Nostoc punctiforme, Anabaena flos-aquae, Spirulina maxima, and Selenastrum capricornutum. Chlorophyll a contents, phycocyanin contents, and photosynthetic activities of the A. flos-aquae decreased evidently in an infected culture for a period. Bacterium B. cereus attacked rapidly A. flos-aquae cells by cell-to-cell contact mechanism. It was shown that the lysis of A. flos-aquae began with the breach of the cyanobacterial cell wall, and the cyanobacterial cell appeared abnormal in the presence of the PL bacterium. Moreover, transmission electron microscope examinations revealed that a close contact between the bacterium and the cyanobacterium was necessary for lysis. Some slime extrusions produced from B. cereus assisted the bacterial cells to be in close association with and lyse the cyanobacterial cells. These findings suggested that this bacterium could play an important role in controlling the Aphanizomenon blooms in freshwaters. (c) 2006 Elsevier Inc. All rights reserved.

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A spin-injection/-detection device has been fabricated based on the multiple quantum well light emitting diode (LED) structure. It is found that only a broad electroluminescence (EL) peak of a full width at half maximum of 8.6 nm appears at the wavelength of 801 nm in EL spectra with a circular luminescence polarization degree of 18%, despite PL spectra always show three well resolved peaks. The kinetic energy gained by injected electrons and holes in their drift along opposite directions broadens the EL peak, and makes three EL peaks converge together. The same process also destroys the injected spin polarization of electrons mainly dominated by the Bir-Aronov-Pikus spin relaxing mechanism.