滇黄芩器官发生与不同继代次数遗传稳定性的RAPD分析

本研究以滇黄芩不带腋芽茎段为外植体,采用不同配比激素的MS培养基建立了滇黄芩器官发生,植株再生体系。结果表明,茎段在MS＋6-BA 2mg／L＋NAA 0.5mg／L培养基上培养时,愈伤组织诱导率达到100％;以相同培养基进行分化,并在MS＋6-BA 2mg／L＋NAA 0.2mg／L培养基上扩繁丛生芽。在1／2MS＋IBA 0.2mg／L培养基诱导生根,扦插继代。利用RAPD分析不同继代次数再生植株,所用的39条随机引物中只有1条引物带型发生变化。说明经组织培养获得的再生植株遗传稳定,可多次继代培养,为滇黄芩进一步遗传操作和扩大药材资源奠定基础。     

发根农杆菌介导转化小金海棠的影响因素

为了研究发根农杆菌介导的小金海棠的遗传转化体系,采用农杆菌侵染的方法,对影响Ri质粒诱导小金海棠产生发状根的各种因素进行了研究,结果发现,Ri质粒转化小金海棠形成发状根的效率,受发根农杆菌种类、浓度、生理状态及小金海棠外植体的种类、外植体在菌液中的侵染时间、共培养时间、基本培养基等多种因素的影响,加入20mg/L AS对转化效率影响不大。采用稀释5倍的处于对数生长期的8196菌株,感染小金海棠无菌苗幼嫩叶片5-10min,在MS培养基上共培养2d后转入含卡那霉素10mg/L、头孢霉素250mg/L的1/4MS诱导培养基上培养一个月,发根诱导效率最高可达93.3%.随机选择50条诱导的发根进行GUS染色,结果发现有96%发根GUS染色呈阳性。对48条GUS阳性的发根进行PCR检测,其阳性率为100%。高效的发根诱导体系为发根的进一步再生及转入外源基因奠定了基础。     

朝天罐的离体培养与植株再生

以朝天罐（Osbeckia opipara）的茎段为外植体进行离体培养植株再生研究,结果表明：茎段在适宜的培养基上可形成愈伤组织,并分化成再生植株。培养基MS + 6-BA 2 mg/L  + NAA 0.2 mg/L + 蔗糖30 mg/L+ 琼脂7 mg/L,既适合于愈伤组织的诱导,也适合于愈伤组织的增殖和分化,诱导率为83.3％;丛生芽增殖的最佳培养基为MS + 6-BA 0.2 mg/L+ NAA 0.1 mg•L-1 + 蔗糖 50 mg/L + 琼脂 7mg/L,增殖系数为 7.2; 试管苗生根的最佳培养基为1/2 MS + NAA 0.5 mg/L + IBA 0.2 mg/L + 6-BA 0.05 mg/L + 蔗糖 20 mg/L + 琼脂 7 mg/L,生根率为100％。     

泡桐体外器官直接发生的植株再生

以毛泡桐、兰考泡桐和白花泡桐的茎段和叶片为材料 ,建立其器官高效直接再生系统。结果表明 ,毛泡桐茎段芽诱导的适宜培养基为MS + 0 1mg LNAA + 1 8mg LBA ,叶片芽诱导的培养基为MS + 0 1mg LNAA + 1 5mg LBA ;兰考泡桐茎段芽诱导的适宜培养基为MS + 0 3mg LNAA + 1 8mg LBA ,叶片芽诱导的适宜培养基为MS + 0 1mg LNAA + 1 5mg LBA ;白花泡桐茎段和叶片芽诱导的最适宜培养基分别为MS + 0 3mg LNAA + 1 8mg LBA和MS + 0 1mg LNAA +1 8mg LBA。此外 ,毛泡桐、白花泡桐和兰考泡桐不同外植体幼芽生根的最适培养基皆为 1 2MS + 0 1mg LNAA。     

水稻巯基蛋白酶抑制剂基因(OCI )转化甘薯获得转基因植株

用根癌农杆菌(Agrobacterium tumefaciens)介导法将水稻巯基蛋白酶抑制剂基因(OCI)导入甘薯（Ipomoea batatas)品种栗子香，获得了转基因植株。所用菌株为根癌农杆菌LBA4404，其携带的pBinh质粒上含有新霉素磷酸转移酶Ⅱ(NPT Ⅱ)基因和OCI基因。将继代培养3d后的栗子香胚性悬浮细胞与LBA4404(OD600nm=0．5)共培养4d。将共培养后的胚性悬浮细胞首先在含有2mg／L2，4-D和300mg／L Carb(carbencillin)、但不含有Kan（kanamycin)的MS液体培养基中培养5d，然后在含有2mg／L2，4-D、50mg／L Kan和300mg／L Carb的MS液体培养基中进行选择培养。选择培养4周后，将直径约1mm的抗Kan细胞团转移到添加2mg／L2，A-D、50mg／L Kan和300mg／L Carb的MS固体培养基上，共转移200个细胞团，诱导得到了8个胚性愈伤组织。将这些抗Kan胚性愈伤组织转移到添加1mg／L ABA、50mg／L Kan和100mg／LCarb的MS固体培养基上，通过体细胞胚胎发生途径获得了13株再生植株。PCR和PCR-Southern检测结果表明，获得的13株再生植株中7株是转基因植株。     

甘薯和近缘野生种Ipomoea triloba的种间体细胞杂种植株再生

摘要: 将甘薯(Ipomoea batatas )品种元气和近缘野生种I. triloba的叶柄原生质体用PEG法融合后, 培养在含有0.05～0.20 mg/L 2,4-D和0.5 mg/L KT的改良MS液体培养基中。培养约7周后，将直径达1～2 mm的小愈伤组织转移到添加0.05～0.20 mg/L 2,4-D和0～0.5 mg/L KT的MS培养基上使其增殖。转移3～5周后，愈伤组织直径达8～15 mm。将这些愈伤组织再转移到MS基本培养基上，或转移到添加 3.0 mg/L BAP的MS培养基上培养4～5周后再进一步转移到MS基本培养基上进行培养，结果获得了69株再生植株。RAPD分析表明，其中1株再生植株是元气和I. triloba的种间体细胞杂种。杂种植株的株型呈野生攀缘型，相似I. triloba；而茎色呈绿-紫色，相似元气；其茎的节间长度和叶片形状表现为融合双亲的中间型。茎尖细胞染色体数为54～103，因细胞不同而不同。     

不同激素处理对山苍子雄性离体再生植株快繁的影响

用大叶山苍子成年雄株带茅茎段为外植体,研究不同激素种类和配比对山苍子离体培养及再生植株快繁的影响效果.结果表明:以MS 6-BA 2.0 mg/L 2.4-天1.0 mg/L培养基适于带茅茎段愈伤组织的诱导分化,分化率达62.5%;以MS 6-BA 2.0 mg/L NAA 0.01 mg/L培养基适于芽苗增殖,增殖倍数为5.3;最佳生根培养基为1/2 MS IAA0.5mg/L IBA0.5mg/L TA2.0MG/L,生根率达83.3%.     

菊苣高效不定芽直接发生及其植株再生

本研究以菊苣无菌苗叶片为外植体,建立了高效不定芽直接发生及其植株再生体系。在附加不同浓度N-6-benzyladenine(6-BA)或与低浓度α-naphthaleneacetic acid(NAA)组合的MS培养基上,5~7d外植体表面不经过愈伤组织诱导阶段,直接形成不定芽。组织学观察表明,不定芽起源于叶片维管束薄壁细胞,且其微管组织系统与叶片外植体内微管组织系统紧密相连。6-BA是不定芽直接发生所必需的,外植体的发育时期、取材部位和培养基蔗糖浓度对不定芽直接发生有重要影响。在附加2.0mg/L 6-BA,0.5mg/L NAA,100mg/L Vc,100mg/L VB1,300mg/L脯氨酸和40g/L蔗糖的MS培养基上,培养20d龄基部叶片15d时,不定芽直接发生频率最高为100%,每块外植体上产生的不定芽数量也最多,平均为36~38个。在1/2 MS+IBA 0.5mg/L培养基上,再生苗诱导生根频率为97.58%,再生植株移栽于盆土中,100%存活且生长良好,未见形态异常。     

尾叶桉茎段再生系统的建立

本文以尾叶桉(Eucalypt urophylla)U6的无菌苗茎段为材料,选用MS培养基为基本培养基,研究不同IAA浓度对尾叶桉茎段愈伤组织诱导,不同IAA、6-BA浓度对尾叶桉茎段愈伤组织诱导芽、生根的影响。结果表明,当外植体的接种数一样,IAA的浓度为20.0mg/L时,愈伤组织诱导率达到最高,为95%,且此浓度下,出芽率、生根率最高,分别达43.3%和100%,而添加0.5mg/L6-BA则会抑制芽的再生、不能产生根。因此最适合诱导尾叶桉愈伤组织、再生芽和再生根的培养体系是MS+20.0mg/L IAA+30g/L蔗糖+8g/L琼脂。该再生系统的建立将为转基因技术改良桉树性状提供前提条件。     

铁皮石斛液体振荡培养的形态学观察

铁皮石斛作为传统的中药材一直被广泛应用。由于铁皮石斛的种子直接萌发非常困难,无菌播种就成为铁皮石斛种苗繁育的主要方法,但不能完全保持母本优良特性。本研究以铁皮石斛茎段无菌腋芽诱导出的类原球茎体（PLBs）为外植体,建立了其液体快速培养技术体系。该体系利用MS＋BA 2 mg/L＋NAA0.1 mg/L培养基,液体振荡培养初期,增殖量为10%-15%（鲜重比）,2个月后增殖量为30%-50%。PLBs最初乳白色,后转变为半透明浅绿色,多数成团,少量为单个;3-4个月,部分PLBs顶部变尖,开始萌发。转接到MS＋BA 0.2 mg/L＋NAA 0.05 mg/L固体培养基上PLBs转为不透明的绿色,并萌发出芽,再转接到MS＋NAA 0.1 mg/L＋香蕉100 g/L固体培养基上壮苗生根,形成健壮的完整植株。该结果可为多种兰花种苗的快速繁育提供直观的参考。     

Transport patterns of foliar and root absorbed copper in bean seedlings

Abstract

Copper absorption by roots or leaf and transport to other parts were followed in 9‐days old bean (Phaseolus vulgaris L. cv. Vaghya) seedlings. Translocation was also measured in 4 cm segments of the stem. It was found that larger amount of Cu was retained in the roots and Cu was more mobile through phloem than through xylem, as indicated by the data on translocation from root and leaf. Bean plants were found to translocate more Cu to the stem than to other parts. Kinetic analysis of absorption by excised roots and stem segments revealed that the roots have a maximum uptake capacity and high affinity for Cu.     

Caffeic acid derivatives production by hairy root cultures of Echinacea purpurea

Inoculation of leaf explants of Echinacea purpurea (Moench) with Agrobacterium rhizogenes induced hairy roots with the capacity to produce biologically active caffeic acid derivatives (CADs), especially cichoric acid. The kinetics of growth, the uptake of macronutrients, and the accumulation of CADs were investigated in heterotrophically cultured hairy roots for a 50 day period. A maximum of 12.2 g L(-1) dry biomass was achieved in MS nutrients supplemented with 30 g L(-1) sucrose on day 40. The mathematical relationship between hairy root growth and conductivity was established during the exponential phase in Erlenmeyer flasks. HPLC analyses of methanolic (0.1% phosphoric acid; 70:30, v/v) extracts from hairy roots revealed the presence of important CADs: cichoric acid (19.21 mg g(-1) dry biomass), caftaric acid (3.56 mg g(-1) dry biomass), and chlorogenic acid (0.93 mg g(-1) dry biomass). These results demonstrate that biotechnological production of CADs in hairy roots of E. purpurea is possible. Furthermore, these hairy root cultures offer, for the very first time, an excellent biological model to study the biosynthetic pathway of medicinally important CADs.     

Inheritance in hexaploid wheat of genes for hairy auricles and hairy leaf sheath derived from Aegilops tauschii Coss.

Inheritance of genes for hairy auricles and hairy leaf sheath of Ae. tauschii in hexaploid wheat backgrounds (synthetic hexaploid wheat and common wheat varieties) was analyzed. The results indicated that hairy auricles and hairy leaf sheath of Ae. tauschii can be transferred and are expressed in hexaploid wheat. In a synthetic hexaploid wheat ('Ae. tauschii' 188) hairy auricles was proved to be controlled by a single dominant gene derived from Ae. tauschii, which was different from the Pa gene located on chromosome 4BS of common wheat. The hairy leaf sheath phenotype of 'Altar 84/Ae. tauschii 188' was also controlled by a single dominant gene derived from Ae. tauschii, which is obviously different from the Hls gene in T. dicoccoides. We suggest to designate the Ae. tauschii genes for hairy auricles and hairy leaf sheath as Pa2 and Hls2, respectively; such genes could be used as useful genetic markers in common wheat.     

玉米毛状根再生植株对水分胁迫的响应

为研究玉米根系对水分胁迫的响应,以玉米毛状根再生植株为材料,在水分胁迫下,测量其生育时期的植株生长和生理指标。结果表明,水分胁迫下玉米毛状根再生植株光合速率、蒸腾速率、细胞间隙CO2浓度、气孔导度均较高。水分胁迫下,毛状根再生植株的根系水导降幅最小,为13.2%,对照品种H99下降了84.7%。各营养器官含水率最高,叶渗透调节能力增强。这说明由于毛状根再生植株强大的根系,保证了植株生长发育过程中的水分供应和光合能力。     

Influence of thigmic stress or chlormequat chloride on tomato morphology and elemental uptake

Root dry weight and leaf number were not affected by thigmic stress or chlormequat chloride. Shoot dry weight, shoot:root dry weight ratio, shoot height, leaf area, and root surface area were decreased by both thigmic stress and chlormequat chloride. However, root length was decreased and root radius increased only by chlormequat chloride. Total element uptake was decreased by both thigmic stress and chlormequat chloride. In the shoot and root, N was not affected, P was increased in the shoots, K decreased in the shoots but increased in the roots, and Mg decreased in the roots as a result of thigmic stress and chlormequat chloride. Whereas, Ca was decreased in the roots and Mg increased in the shoots by chlormequat chloride. The uptake of N, P, Ca, or Mg was not affected by thigmic stress or chlormequat chloride, however, K uptake per root surface area decreased. Thigmic stress decreased K in both the upper and lower stem but did not affect leaf concentration of K. However, chlormequat chloride decreased K in both the stem and leaves. Thigmic stress because of the role of K in cell elongation. Chemical name used: (2‐chloroethyl)trimethyl‐ammonium chloride (chlormequat chloride).     

影响人参发根皂苷含量基因rolC的克隆与序列分析

摘要：通过发根农杆菌?穴Agrobacterium rhizogenes ?雪与人参?穴Panax ginseng C. A. Mey?雪根外植体共培养，用直接接菌方法诱导出人参发根。培养4周后人参发根的总皂苷含量达到栽培3年生人参皂苷含量的水平，单体皂苷Rb1含量明显提高，说明TL-DNA具有影响人参皂苷生物合成的能力。根据Slightom等RiA4TL-DNA序列分析结果，利用PCR方法从人参发根中扩增并克隆了影响人参皂苷合成的基因rolC。与已发表序列相比较，核苷酸序列的同源性为99.9%。     

小麦根系物质流示踪研究

在小麦生育进程各主要叶龄期分别用~(32)P和~(14)C标记各根层和叶层,研究根系营养物质流与地上部光合同化物质流的动态变化及兵在产量形成上的相互关联作用。研究表明,小麦各层次生根的最大功能同主茎叶龄进程基本上保持同步关系,即n/0叶龄发生的根至n+3/0叶龄发挥显著吸收功能。小麦某一叶龄期的功能叶和功能根在同化物质和营养物质输配上表现出互相依赖、相互促进的关系。     

~(60)Coγ射线辐照对广藿香离体培养的影响

研究60Coγ射线辐照对广藿香离体培养的影响,为将辐照诱变技术应用于广藿香新品种选育奠定基础。以广藿香的叶片、带节茎、不带节茎及根为材料进行60Coγ射线辐照试验,采用MT基本培养基,附加0.05mg/L BA进行离体培养。广藿香外植体的死亡率随着辐照剂量的提高而上升,由回归方程导出广藿香叶片、带节茎及不带节茎的辐照半致死剂量分别为72、66和64Gy;外植体再生芽能力,随着辐照剂量的升高而降低。表明辐照会造成外植体的损伤及死亡,对外植体的离体再生有明显的抑制作用,同时,也使部分外植体的再生苗出现一些表型的变化。     

不同辣椒品种镉吸收积累能力及关键期研究

  【目的】   比较不同辣椒品种植株Cd的吸收和积累动态差异,明确Cd积累关键期,为生产上规避辣椒Cd积累提供技术支持。   【方法】   以8个辣椒品种为材料进行了田间小区试验,供试土壤全镉含量为0.194 mg/kg。在辣椒苗期 (移栽当天)、壮苗期 (移栽后32 天)、结果期 (移栽后65 天)、成熟期 (移栽后111天) 采样,分析辣椒植株根、茎、叶及果实生物量、Cd含量,计算不同组织间的Cd运转系数。   【结果】   供试8个品种分为菜椒和朝天椒两类。4次采集的样品,叶部Cd含量均为菜椒 > 朝天椒,果实Cd含量在结果期菜椒高于朝天椒,成熟期朝天椒显著高于菜椒。朝天椒果实以品种艳椒425的Cd含量最低,菜椒以品种苏椒5号Cd含量最低。壮苗期为辣椒植株Cd的快速积累期,朝天椒根、茎和叶Cd含量移栽后0到32天分别增加了14.9、51.2和51.9倍,菜椒分别增加了8.9、25.6和39.9倍。结果期Cd积累变缓,移栽后65天朝天椒根、茎和叶Cd含量比移栽后32 天分别增加了0.35、0.55和–0.13倍,菜椒分别增加了0.20、0.01和–0.29倍;成熟期 (移栽后111 天) 朝天椒根、茎和叶Cd含量比结果初期 (移栽后65天) 分别增加了–0.29、0.44和–0.40倍,菜椒分别增加了–0.34、–0.14和–0.32倍。不同辣椒品种Cd在植株内的转运能力有差异,4次采样根–茎Cd转运效率菜椒 > 朝天椒,而茎–叶、叶–果和茎–果Cd转运效率则是朝天椒 > 菜椒。辣椒果实Cd积累量与壮苗期茎的Cd含量呈极显著正相关 (P < 0.01),与茎–果、叶–果Cd转运效率显著相关 (P < 0.05)。   【结论】   壮苗期前 (移栽后0～32天) 为辣椒植株Cd的快速积累期,该时期辣椒根茎叶中的Cd含量可以增加数十倍,进入结果期Cd含量的增加速率大大降低。壮苗期辣椒茎的Cd含量与果实Cd积累量呈极显著相关。菜椒根中的Cd向茎的转运能力较强,而朝天椒茎叶中的Cd向果实中转运能力较强,因而,朝天椒果实Cd积累能力高于菜椒。