Stabilization of double-stranded oligonucleotides using backbone-linked disulfide bridges.

A convenient, practical route to the synthesis of disulfide-bridged oligonucleotides has been developed. Aliphatic linkers with terminal thiol groups have been attached to the phosphodiester backbones of partially or fully complementary oligonucleotide sequences and oxidized to yield covalently closed oligonucleotides with disulfide bridges. This procedure has been used to prepare a duplex with disulfide bridges at both ends and stem-loop sequences with single disulfide bridges. Oxidation of a self-complementary duplex possessing terminal thiol groups produced both hairpin and duplex structures with disulfide bridges, the relative proportions of each being dependent upon the reaction conditions. These bridged hairpin and duplex structures were shown to be interconvertible by reduction and re-oxidation. The melting profiles of disulfide-bridged oligonucleotides were compared with the same sequences without bridges and with sequences possessing triethylene glycol bridges, and in all cases the introduction of disulfide bridges resulted in a considerable increase in thermal stability. EcoRI endonuclease was capable of cleaving a disulfide-bridged duplex possessing a recognition site for this enzyme, thus supporting a lack of distortion of the recognition site. The disulfide bridges could be cleaved using a large excess of DTT to regenerate the corresponding sulfhydryl compounds. A study of the serum stabilities of disulfide-bridged oligonucleotides showed that the bridged duplexes were much more stable than their unmodified counterparts, whereas the rate of degradation of the stem-loop structures was more dependent upon the size of the loop than the presence or absence of the disulfide bridge. In summary, we have described a novel methodology, employing commercially available reagents, for the stabilization of oligonucleotide duplexes or stem-loop structures by disulfide bridge formation.     

Discrete differences between strains of different Rhizobium spp. for competitive nodule occupancy on beans

Rhizobium etli strain TAL182 and R. leguminosarum bv phaseoli strain 8002, both of which produce melanin pigment, were tested for their nodulation competitiveness on beans by paired inoculation with two strains which do not produce melanin: R. tropici strain CIAT899 and Rhizobium sp. strain TAL1145. An assay was developed to distinguish nodules formed by the melanin-producing and non-producing strains. Strain TAL182 had discrete competitive superiority over CIAT899 and TAL1145 for nodulation of beans. Nodulation competitiveness was not correlated with the ability to produce melanin pigment or the host range of the Rhizobium strains tested.The authors are with the Department of Plant Molecular Physiology, University of Hawaii, 3050 Maile Way, Gillmore 402, Honolulu, HI 96822, USA     

提高赤红壤旱地生态系统土壤肥力问题的研究

提高赤红壤旱地生态系统土壤肥力问题的研究高志强（福建农业大学土地与环境学系,福州３５０００２）ＳｏｉｌＦｅｒｔｉｌｉｔｙＩｍｐｒｏｖｅｍｅｎｔｏｆＵｐｌａｎｄＬａｔｅｒｉｔｉｃＲｅｄＳｏｉｌＥｃｏｓｙｓｔｅｍ．￥ＧａｏＺｈｉｑｉａｎｇ（Ｄｅｐａｒｔｍ...     

中国生态环境区划初探

中国生态环境区划初探高密来（中国人民大学区域经济研究所北京１００８７２）ＡｎＡｐｐｒｏａｃｈｔｏｔｈｅＲｅｇｉｏｎａｌｉｚａｔｉｏｎｏｆＥｃｏｌｏｇｉｃａｌＥｎｖｉｒｏｎｍｅｎｔｉｎＣｈｉｎａ．￥ＧａｏＭｉｌａｉ（ＩｎｓｔｉｔｕｔｅｏｆＲｅｇｉｏｎａ...     

买麻藤茎的结构及其异常次生生长

买麻藤(Gnetum montanum Mgr.)茎的各类组织的排列与被子植物的茎非常相似,其次生生长也属同一类型。其中最明显的相似之处,是木质部中的导管和韧皮部中的筛分子与“伴胞”。藤本植物买麻藤,正常的次生生长开始后,由每一维管束外侧的薄壁组织经脱分化产生新的形成层,以后逐渐形成一圈维管束。这与前人所描述的新形成层来自韧皮薄壁组织是不同的。异常的维管束与正常的一样,由木质部和韧皮部组成,并被髓射线分隔呈楔形。当第一轮维管束停止生长以后,在其外围以同样方式形成新的一轮,以后并可连续形成多轮。     

羧基化多壁碳纳米管、混合盐及其复合胁迫对水稻幼苗生理特性的影响

将水稻（Oryza sativa L.）幼苗悬浮培养于含有羧基化多壁碳纳米管MWCNTs-COOH（0、2.5、5.0、10.0 mg/L）、50 mmol/L混合盐（1NaCl：9Na₂SO₄：9NaHCO₃：1Na₂CO₃）,以及MWCNTs-COOH+混合盐的复合溶液中,10 d后检测叶片生理生化指标变化,研究MWCNTs-COOH复合盐碱胁迫对水稻幼苗的毒性及生态风险。结果显示,与对照组相比,MWCNTs-COOH单一组诱导下水稻叶片O₂^·-和H₂O₂的产生不明显,而混合盐组和混合盐+MWCNTs-COOH复合组均诱导了O₂^·-和H₂O₂产物的大量累积。MWCNTs-COOH与混合盐复合后,加剧了O₂^·-和H₂O₂的累积,并有明显的浓度效应。活性氧（ROS）作为信号分子在一定程度上诱导了各处理组部分抗氧化酶（SOD、CAT、POD、APX）活性的升高;与混合盐组相比,低浓度混合盐+MWCNTs-COOH复合组中叶绿素a和胡萝卜素含量呈一定程度的升高;MWCNTs-COOH与混合盐复合后,抑制了叶片中可溶性糖（SS）和脯氨酸（Pro）的合成,致使相对电导率（REC）和丙二醛（MDA）含量显著升高。上述抗氧化酶活性及叶绿素a和胡萝卜素含量的升高对缓解水稻叶片氧化损伤、维持正常的光合电子传递及对过剩光能的热耗散是有益的,是水稻幼苗重要的防御机制。本研究表明MWCNTs-COOH单一处理在一定程度上诱导了水稻叶片的氧化胁迫和应激响应,与混合盐复合后加剧了叶片的氧化胁迫和应激损伤。     

荧光显微镜观察大蒜油对腹水癌细胞的影响

用吖啶橙染色,荧光显微镜观察大蒜油对—S180,昆明小鼠,U14 C57BL/6J小鼠和L1210DBA小鼠的癌细胞作用的影响,结果表明大蒜油有直接破坏癌细胞的DNA,RNA和分裂中期染色体的作用。给药后2—6小时作用最强,12小时后癌细胞数逐渐恢复,因此给药间隔不应超过12小时。我们研究证实大蒜油有明显的抗癌作用,为临床应用提供有力的依据。     

浙北平原水杉人工林生物量的研究

 本文对浙江省北部平原水杉人工林的生物量和生产力进行了测定和研究。按平均标准木法分别调查了干、枝、叶、根的生物量。据调查材料,用相对生长规律建立了估测单株林木各器官干重的回归方程,方程的相关系数和估测精度符合要求,具实用价值。研究结果表明：水杉人工林生物量随年龄的增长而增加,林带18年生后增加速度变缓,片林生物量普遍大于林带生物量,生物量组成比例因年龄而异。随着年龄的增加,年平均净生产量、叶面积指数、光能利用率均逐步加大,至18年生时,上述指标分别为17.51t／ha·a、9.1和0.77%。叶净光合生产率以速生阶段为最大,衰退阶段为最低,当叶净光合生产率急剧下降时,可实施主伐更新。