Molecular Cloning and Sequence Analysis of DNA in Mycoplasma-Like Organism of the Paulownia Witches Broom

Partially purified mycoplasma-like organism (MLO) preparations were obtained from diseased paulownia with witches' broom (PWB). EcoR Ⅰ -Hind Ⅲ digested fragment of the total DNAs extracted from the preparations was Iigated into pGEM-3Zf (+) vector. The recombinant molecules were transformed into Escherichia coli strain DH5α. After screening by differential hybridization and identification with Dot and Southern blot hybridization analysis, two clones (A4, 1.69 kb and C42, 2.08 kb) were obtained. Their DNA inserts as probes hybridized only with total DNAs from PWB-diseased plants but not with extracts from healthy plants. Abundant (A+T) content was found in the sequences of DNA inserts from the two clones, 72.5% (A4) and 67.9% (C42, respectively, which possessed the character of DNA from mycoplasma spp.     

Production and Preliminary Application of Monoclonal Antibodies Against Paulownia Witches' Broom MLO

Monoclonal antibodies against mycoplasma-like organisms (MLO) associated with paulownia witches’broom (PWB) were produced by using partially purified preparations from diseased paulownia. Splenic cells from immunized mice were fused with sp2/0murine myeloma cells. Screened by indirect ELISA using partially purified PWB-MLO and healthy paulownia extracts as detecting antigens, two hybridoma clones that stably secreted specific antibodies against PWB-MLO were obtained from 459 clones of four successful fusions. The monoclonal antibodies were isotyped and determined to be immunoglubin classes IgG2a and IgG3. Antibody titers of ascitic fluids were both over 1.6 × 104 assayed by indirect ELISA. Priliminary application on several specimens proved that they were the monoclonal antibodies against PWB-MLO.     

白花泡桐不定根发生过程中内源激素和RNA的变化

白花泡桐（Ｐａｕｌｏｗｎｉａｆｏｒｔｕｎｅｉ（Ｓｅｅｍ．）Ｈｅｍｓｌ．）成年型和幼年型茎切段体外培养时不定根发生过程中内源ＩＡＡ、ＣＴＫ和ＡＢＡ含量测定表明：幼年型材料中内源ＩＡＡ和ＣＴＫ的含量在诱导的第２天同时达到高峰,而成年型材料中ＩＡＡ和ＣＴＫ含量的高峰则在第４天出现．两种类型切段根原基出现的时间都与其内源ＩＡＡ和ＣＴＫ的高峰一致．幼年型材料的内源ＡＢＡ含量在第４天达到高峰,随后迅速下降．成年型材料中内源ＡＢＡ则逐步下降．成年型和幼年型材料中ＲＮＡ的变化相同,在诱导的第２天稍有下降,随后显著增加．结果显示,不定根的发生与其内源激素和ＲＮＡ的变化密切相关．     

High Frequency Plant Regeneration from Nodal Explants of Paulownia elongata

Abstract: High frequency of plant regeneration from Paulownia elongata was obtained on Murashige and Skoog (MS) medium and Woody Plant Medium (WPM), with appropriate supplements of growth regulators. Leaves, leaves with petioles, internodes and nodes excised from 3-month-old non-aseptically grown P. elongata were used as explants. The highest shoot regeneration efficiency (93.7 %) was obtained from the nodes of P. elongata on MS medium supplemented with 0.1 mg/ml naphthaleneacetic acid (NAA) and 1 mg/ml 6-benzylaminopurine (BAP). The highest root formation efficiency (100 %) from the regenerated shoots was obtained on WPM supplemented with 1 mg/ml indolebutyric acid (IBA). Rooted plantlets were transplanted to soil with a survival efficiency of almost 100 %. The regeneration system reported here could be useful for rapid multiplication of elite genotypes of P. elongata in a short period of time.     

安徽涡阳县桐农间作类型及其效益分析

一、概况涡阳县位于黄淮平原南部,安徽省的西北部。全县土地面积2.09×10~5ha,其中耕地面积1.44×10~5ha。人口105万。本县属暖温带半湿润季风气候区,年平均气温14.6℃,降水量824mm,蒸发量1974.5mm。太阳辐射总量5.22×10~5J·m~(-2)。气候温和,水热同季,     

不同种源白花泡桐种子的休眠生理生态研究

为了探讨亚热带树木休眠理论以及休眠与树木分布的相互关系,研究了不同种源白花泡桐种子的休眠生理生态学特性。结果表明：白花泡桐种子适宜发芽温度范围在20～30℃,经过低温处理后,适宜发芽温度范围扩大到15～30℃,具有浅低温休眠特性;5℃的低温处理不仅提高了不同种源白花泡桐种子的发芽率和发芽势,而且加快了发芽速度,尤其在15℃发芽温度条件下的促进效果更加明显;40d的5℃低温处理效果最好．20d、60d的低温处理效果相对较差;没有用低温处理的不同种源白花泡桐种子,除了在25℃发芽温度下的发芽势差异不显著外,在不同发芽温度条件下的发芽率与发芽势都存在着显著差异,但在20℃、25℃发芽温度下的平均发芽时间差异不显著。在15℃发芽温度条件下,除了桂林种源以外,没有用低温处理的白花泡桐种子的发芽率与发芽势都随着种源纬度的增加而增加,而在20～30℃的适宜发芽温度范围内随种源纬度的变化趋势不明显;不同种源白花泡桐种子对低温处理反应差异明显。低温处理后的中亚热带以南的低纬度种源白花泡桐种子发芽率、发芽势都较北亚热带的高纬度种源高。低温处理后的中亚热带株州种源和北亚热带南部的宜昌种源白花泡桐种子的平均发芽时间最长,而随着纬度的降低或增加平均发芽时间都变短。不同种源白花泡桐种子的浅休眠特性不同,是对不同种源地气候的一种生理生态适应。     

根癌农杆菌对健康和患丛枝病泡桐的遗传转化

选取健康及患丛枝病泡桐（Paulownia spp.)为材料,建立组织培养和植株再生系统,以茎段作为转化受体,诱导分化和生根的最佳激素组合分别是MS＋BA4mg/L NAA0.2mg/L和1／2MS＋KT0.5mg/L IBA0.25mg/L。芽分化频率可达22．8％。健康和患病泡桐的茎段经农杆菌共培养3d后,在附加50mg/Lm的选择分化培养基上培养20d左右再生出抗性芽,经培养、诱导生根,获得了转基因再生植株,建立了泡桐的遗传转化体系。PCR和Southern杂交检测证明外源基因已整合到泡桐的基因组,标记基因ITPⅡ在再生植株中也得到表达,同时对影响转化的一因素进行了探讨。     

泡桐叶片蛋白质多态性及其聚类分析

研究了9种泡桐叶片蛋白质的多态性,并根据其叶片蛋白质聚丙烯酰胺凝胶单向和双向电泳结果,将它们聚类为白花泡桐组[白花泡桐（Paulownia fortunei）和白花兰泡桐（P.elongata f.allba）]、南方泡桐组[南方泡桐（P.australis )和成都泡桐（P.albiphloea var.chengtuensis)]和毛泡桐组（毛泡桐（P.tomentosa）、川泡桐（P.fargesii）、鄂川泡桐（P.albiphloea）、山明泡桐（P.lamprophylla）和兰考泡桐（P.elon-gata）]。该结果可为了解泡桐属植物的亲缘关系和种的鉴定提供参考依据。     

泡桐愈伤组织再生植株的诱导与培养

首先以MS为基本培养基从 1 8个不同浓度的NAA和BA组合中 ,找出了毛泡桐 (Paulowniatomen tosa)、南方泡桐 (Pauiowniaaustralis)、白花泡桐 (Paulowniafortunei)、兰考泡桐 (Paulowniaelongata)和豫杂一号泡桐 (Paulowniatomentosa×P .fortunei)叶片愈伤组织诱导芽分化最适培养基分别为MS 0 .3NAA 1 2BA、MS 0 .3NAA 1 2BA、MS 0 .5NAA 1 2BA、MS 0 .5NAA 1 2BA和MS 0 .7NAA 1 2BA ;然后 ,筛选出了 5种泡桐芽诱导根的最适培养基 (分别为 1 /2MS 0 .1NAA、1 /2MS 0 .1NAA、1 /2MS、1 /2MS 0 .3NAA和 1 /2MS 0 .5NAA)。这些结果为利用不同种泡桐的原生质体融合培育泡桐新品种奠定了一定的基础。