氯虫苯甲酰胺对苹果树桃小食心虫及金纹细蛾的控制作用

2006～2007年,对35%氯虫苯甲酰胺水分散粒剂进行田间药效试验。结果表明,35%氯虫苯甲酰胺水分散粒剂是防治苹果树桃小食心虫Carposina niponensisi Walsingham和金纹细蛾Lithocolletis ringoniella Mats的有效药剂。其有效成分为35～50mg/kg处理防治桃小食心虫,药后5d的防效在83%～100%之间,药后35d防效在77%以上,整体防效不及对照药剂4.5%高效氯氰菊酯乳油30mg/kg处理;10～20mg/kg处理防治金纹细蛾,药后5～15d防效在58%～100%之间,各期防效优于生产常用药剂25%灭幼脲悬浮剂125mg/kg处理。     

虫酰肼和甲维盐对甜玉米草地贪夜蛾的生物活性及田间防效

为了研究虫酰肼和甲氨基阿维菌素苯甲酸盐(甲维盐)对新入侵我国的草地贪夜蛾毒杀活性并为田间应用提供依据,采用浸叶法进行了室内毒力测定,采用喷雾法进行盆栽试验和田间小区试验。毒力测定表明虫酰肼对草地贪夜蛾2龄、3龄幼虫48 h LC_(50)分别为0.30 mg/L和0.26 mg/L,甲维盐对草地贪夜蛾2龄、3龄幼虫48 h LC_(50)分别为0.005 mg/L和0.002 mg/L,其LC_(50)活性是虫酰肼的60～128倍。盆栽试验表明,20%虫酰肼SC在20 mg/L、5%甲维盐ME 1mg/L喷雾处理后7 d后防效均为100%,在甜玉米田间小区试验最高防效均可达到90%以上。本研究结果表明,虫酰肼可推荐用于草地贪夜蛾的防治并可与甲维盐轮换使用或复配使用。     

氯虫苯甲酰胺对草地贪夜蛾的毒力及田间防效

草地贪夜蛾Spodoptera frugiperda(Smith)是威胁粮食安全的重要害虫,本文测定了氯虫苯甲酰胺对草地贪夜蛾的室内毒力及田间防效,从中筛选出高效的施药方法和施药剂量,为当前草地贪夜蛾防治提供合理用药的参考。结果表明,在室内条件下,使用氯虫苯甲酰胺直接喷雾处理对草地贪夜蛾3龄和4龄幼虫毒力最高,LC_(50)值分别为0.42 mg/L和0.57 mg/L,使用0.5 mg/L 20%氯虫苯甲酰胺悬浮剂对草地贪夜蛾幼虫毒力较高,在处理后第5天均有超过90%的死亡率。在田间条件下使用氯虫苯甲酰胺对草地贪夜蛾幼虫有较好的防效,使用60.0 g.a.i./ha 0.4%氯虫苯甲酰胺颗粒剂喇叭口点施防治低龄幼虫防效较好,药后第1天、第3天和第7天低龄幼虫(1～3龄)的防效分别为90.3%、100%和100%,使用22.5 g.a.i./ha 20%氯虫苯甲酰胺悬浮剂喇叭口喷雾防治高龄幼虫(4～6龄)的防效较好,药后第1天、第3天和第7天高龄幼虫的防效分别为71.7%、84.5%和95.3%。     

四霉素对茶藨子葡萄座腔菌Botryosphaeria ribis室内毒力测定及其田间防效试验

为确定四霉素对杨树溃疡病的防治效果,采用菌丝生长速率法测定四霉素水剂对茶藨子葡萄座腔菌Botryosphaeria ribis的室内毒力,采用刮治喷涂法对茶藨子葡萄座腔菌引起的杨树溃疡病进行田间应用效果研究。试验结果表明,四霉素对茶藨子葡萄座腔菌的EC_(50)和EC_(90)分别为114.82 a.i.mg/L、389.05 a.i.mg/L,对茶藨子葡萄座腔菌具有较好的抑菌效果,优于对照药剂多抗霉素。田间防效试验结果表明,刮治喷涂0.3%四霉素水剂可以有效防治真菌病原茶藨子葡萄座腔菌导致的杨树溃疡病,其有效成分浓度为60～150 a.i.mg/L时,田间防效达68.25%～82.41%,与室内毒力测定结果相符。     

喷雾方式及喷液量对吡蚜酮和啶虫脒在棉田的沉积分布及棉蚜防治效果的影响

为探明防治棉田棉蚜Aphis gossypii (Glover)的最佳喷雾方式及喷液量, 提高棉田的农药利用率, 作者于2011-2012年在山东省棉花苗期和成株期分别使用背负式手动喷雾器和背负式机动弥雾机以常规大容量和低容量喷雾, 比较杀虫剂25%吡蚜酮可湿性粉剂和3%啶虫脒乳油的喷雾雾滴在棉花田的沉积分布及棉蚜防治效果。结果表明： 在棉花苗期, 3%啶虫脒乳油用量450 mL/ha时, 使用机动弥雾机以75, 150和225 L/ha喷液量喷雾, 药剂在地面的流失率分别为24.4%, 28.9%和26.7%; 使用手动喷雾器以300, 450和600 L/ha喷液量喷雾, 杀虫剂在地面的流失率分别为35.6%, 37.8%和46.7%; 啶虫脒不同喷雾处理对棉蚜的防效无显著性差异（P>0.05）。在棉花成株期, 25%吡蚜酮可湿性粉剂用量为300 g/ha时, 使用手动喷雾器以600 L/ha喷液量喷雾, 药剂地面流失率为13.3%; 使用机动弥雾机以喷液量150 L/ha喷雾时, 药剂地面流失率为3.3%; 25%吡蚜酮可湿性粉剂用量减少至225 g/ha, 使用机动弥雾机以喷液量150和300 L/ha喷雾, 对棉蚜的防效与吡蚜酮用量300 g/ha、 使用手动喷雾器在喷液量600 L/ha条件下喷雾相比没有显著差异（P>0.05）。使用机动弥雾机喷雾可以减少田间用药量和喷液量, 降低药液的流失率, 减轻对环境的污染。     

不同杀虫剂对西花蓟马的室内毒力及田间药效

【目的】西花蓟马是一种世界性的危险性入侵害虫,筛选防治西花蓟马的有效药剂可为生产中科学用药提供依据。【方法】通过室内毒力测定和田间药效试验,测定了2种化学药剂和8种生物药剂对西花蓟马成虫的致死中浓度(LC_(50))和药后不同时期的防治效果。【结果】室内毒力测定依据LC_(50)值将各药剂对西花蓟马的敏感性由高到低依次排序为乙基多杀菌素(0.1958 mg·L~(-1))、印楝素(0.9399 mg·L~(-1))、苦参碱(1.2483 mg·L~(-1))、阿维菌素(1.8096 mg·L~(-1))、高效氯氰菊酯(4.4458 mg·L~(-1))、藜芦碱(10.7628 mg·L~(-1))、鱼藤酮(18.1898 mg·L~(-1))、吡虫啉(46.3964 mg·L~(-1))、松脂酸钠(131.5214 mg·L~(-1))、苏云金杆菌(446.2318 mg·L~(-1));田间药效试验发现,乙基多杀菌素和吡虫啉防治西花蓟马均表现出较强的速效性和持效性,药后1～14 d的防效分别可达84%和73%以上;其次是藜芦碱,药后1～14 d的防效可达48.15%～61.37%;高效氯氰菊酯的防效较低,药后14 d的防效为46.22%;生物药剂阿维菌素、苏云金杆菌、苦参碱、印楝素的速效性均较低,但防效随施药后时间的延长而逐渐上升;鱼藤酮持效性最低。【结论】乙基多杀菌素和吡虫啉可推荐为生产中防治西花蓟马的首选药剂,植物源药剂藜芦碱和生物药剂阿维菌素可作为交替使用药剂。     

几种药剂及施药方式对稻纵卷叶螟的防治效果

对稻纵卷叶螟Cnaphalocrocis medinalis Guen e高龄幼虫防治药剂31%氟虫腈.三唑磷ME和当地几种常用药剂及不同施药方法对防治效果影响进行试验。结果表明:(1)毒死蜱(48%乐斯本EC)有较好速效性,药后1 d可达理想防效,三唑磷与氟出腈、氟铃脲复配的药剂(31%氟虫腈.三唑磷ME、21%氟虫腈.三唑磷EC、15%氟铃脲.三唑磷EC)药后3d才能达到理想防效;(2)药后3d,各处理均达到理想防效,防效依次为31%氟虫腈.三唑磷ME1.5L/hm2>31%氟虫腈.三唑磷ME1.05L/hm2>21%氟虫腈.三唑磷EC1.5L/hm2>48%毒死蜱EC1.5L/hm2>15%氟虫腈.三唑磷EC1.5L/hm2>15%阿维菌素.毒死蜱EC1.5L/hm2;(3)分虫龄防效统计,药后3d供试药剂对1龄以下(施药时虫龄,后同)幼虫防效均在91%以上,对2~3龄幼虫,31%氟虫腈.三唑磷ME保持90%以上的防效,其它处理防效明显下降,对4龄以上高龄幼虫,31%氟虫腈.三唑磷1.5L/hm2细喷雾处理防效仍达95.6%,其它处理防效很低。(4)细喷雾可提高31%氟虫腈.三唑磷ME对稻纵卷叶螟的防治效果,尤其是对高龄幼虫,与工农16型1.8mm孔径喷片的常规喷雾相比,1.2mm孔径喷片细喷雾处理药后3 d防效提高5.5%,东方红18型弥雾处理防效提高7.8%,其中,弥雾处理对1~2龄幼虫防效提高4.7%,对4~5龄幼虫防效提高了20.8%。     

毒死蜱与阿维菌素增效混配制剂对斑潜蝇属害虫的防治效果

通过有机磷杀虫剂毒死蜱与生物源农药阿维菌素混配制剂对美洲斑潜蝇Liriomyza sativae室内毒力实验,测定共毒系数为165～234,处于明显增效范围内。据此确定最佳配比和次佳配比,配制该增效混剂30%渗透型可湿性粉剂-1和-02,在山东防治美洲斑潜蝇幼虫的田间试验表明药效优良。制剂用量50 g/667m²药后3,7,11天,两可湿粉的校正防效分别为90.43%～91.71%和87.09%～90.53%,可湿粉-1用量25 g/667m²防效为85.96%～88.28%,可湿粉-2用量37.5 g/667m²相应校正防效为84.01%～85.38%,两增效混剂防治斑潜蝇速效性和持效性皆佳,成本有所下降,且对南美洲斑潜蝇L. huidobrensis亦有较好防效。使用可湿粉与乳油相比较可减少投放入环境的化学品数量。     

防治甘草萤叶甲生物源农药筛选及其对生物多样性的影响

为了筛选出防治甘草萤叶甲Diorhbda tarsalis Weise的生物药剂,采用多种生物源药剂,通过室内、田间试验,进行综合分析。室内毒力测定结果表明,甘草萤叶甲对0.5%黎芦碱SL的敏感性最高,LD50为0.139mg/L,0.3%印楝素EC、L2、1%苦参碱SL、L1依次递减,LD50分别为0.457,1.352,2.014和2.251mg/L,均高于其他药剂。田间防效结果表明,黎芦碱药后21d对甘草萤叶甲的防效最高,为100%,苦参碱、印楝素次之,均为86.67%,L2、L1的分别为66.67%和40%;对非靶标害虫小绿叶蝉也有较好的控制作用,药后21d防效L1较低,为56.67%,其他防效均高于68.89%;对田间天敌多异瓢虫和中华草蛉的安全性,除黎芦碱外,印楝素、苦参碱、L2、L1均较好,药后21d校正虫口减退率最高为57.14%;另外,苦参碱药后各期生物多样性指数平均最高,为2.93,印楝素次之,为2.88,藜芦碱、L1、L2依次递减,分别为2.45,2.43和2.07。因此,印楝素、苦参碱是防治甘草萤叶甲的理想药剂。     

枯草芽孢杆菌菌剂对人参黑斑病的生物防治效果

为明确枯草芽孢杆菌可湿性粉剂(WP)对人参黑斑病的防治效果及对人参生长的安全性,采用生长速率法测定枯草芽孢杆菌对人参黑斑病菌的室内抑菌效果,采用田间小区试验评价枯草芽孢杆菌对人参黑斑病的防治效果及其对人参的安全性。室内抑菌试验结果表明:枯草芽孢杆菌对人参黑斑病菌的菌丝生长具有较好的抑制作用,EC_(50)和EC_(90)分别为0.690 mg/L和103.402 mg/L。两年三地田间试验结果表明:1 000亿CFU/g枯草芽孢杆菌WP在3种施用剂量下对人参黑斑病均有较好的防治效果,平均防效在69.02%～80.95%,其中使用剂量为80 g/667 m~2时防效最好,与对照药剂250 g/L嘧菌酯SC防效差异不显著(P0.05)。该药剂对人参生长无不良影响。1 000亿CFU/g枯草芽孢杆菌WP对人参安全,可用于人参黑斑病的防治,建议在人参发病前或发病初期茎叶喷雾,推荐剂量为60～80 g/667 m~2。     

丹参联合β-榄香烯对肝星形细胞LX-2增殖和凋亡的影响

目的：探讨丹参联合B-榄香烯对肝星形细胞LX-2增殖和凋亡的影响。方法：体外培养肝星形细胞LX-2,分别将丹参（浓度为1、2、4、6、8mg／ml）,β-榄香烯（浓度为25、50、100、150、200μg／ml）,单独作用于LX-2细胞后24h、48h用CCK-8（CellCountKit-8）法检测细胞的增殖情况,并选取合适的药物浓度（丹参3．6mg／ml,β-榄香烯125μg／ml）,然后进行联合用药,加药24h、48h后用CCK-8（Cell CountKit-8）法检测细胞的增殖情况,用流式细胞术检测细胞的凋亡率。结果：OCCK-8法显示丹参（浓度为1、2、4、6、8mg／ml）,8-榄香烯（浓度为25、50、100、150、200μg／ml）作用LX-2细胞24h、48h后,其增殖抑制率均明显高于正常对照组（P〈0．05）,并且呈浓度和时间依赖性。②联合用药（丹参3．6mg／ml,β-榄香烯125μg／ml）时,LX-2细胞的增殖抑制率和凋亡率均显著高于单独用药（P〈0．01）。结论：丹参、β-榄香烯单独或二者联合作用均能抑制LX-2细胞的增殖,且联合应用的作用显著高于单独用药,可协同促进LX-2细胞的凋亡。     

Down-regulation of FoxO-dependent c-FLIP expression mediates TRAIL-induced apoptosis in activated hepatic stellate cells

Activated hepatic stellate cells which contribute to liver fibrosis have represented an important target for antifibrotic therapy. In this study, we found that TRAIL inhibited PI3K/Akt-dependent FoxO phosphorylation and relocated FoxO proteins into the nucleus from the cytosol in activated human hepatic stellate LX-2 cells. The accumulated FoxO proteins in the nucleus led to down-regulation of c-FLIP_L/S expression, resulting in the activation of apoptosis-related signaling molecules including the activation of caspase-8, -3, and Bid, as well as mitochondrial cytochrome c release. These results were supported by showing that siRNA-mediated knockdown of FoxO led to restoration of c-FLIP_L/S expression and resistance to TRAIL-induced apoptosis after treatment of LX-2 cells with TRAIL. Furthermore, c-FLIP_L/S-transfected LX-2 cells showed the decreased sensitivity to TRAIL-induced apoptosis. Collectively, our data suggest that sequential activation of FoxO proteins under conditions of suppressed PI3K/Akt signaling by TRAIL can down-regulate c-FLIP_L/S, consequently promoting TRAIL-induced apoptosis in LX-2 cells. Therefore, the present study suggests TRAIL may be an effective strategy for antifibrotic therapy in liver fibrosis.     

Purification and characterization of large and small subunits of ribulose 1,5-bisphosphate carboxylase expressed separately in Escherichia coli

Procedures were developed for 95 and 80% purification to homogeneity of the large subunit (L) and small subunit (S) of ribulose 1,5-bisphosphate carboxylase/oxygenase (L8S8) from Synechococcus PCC 6301, each expressed separately in Escherichia coli. Purified L had a low specific activity in the absence of S (0.075 mumol CO2 fixed/mg holoenzyme/min). Following elution on a Pharmacia Superose 6 or 12 gel filtration column, 50% of the purified L appeared as the octamer, L8. The rest was in equilibrium with lower polymeric species and/or was retained on the column. Large and small subunits assembled rapidly into the L8S8 holoenzyme that had high specific activities, 6.2 and 3.1 mumol CO2 fixed/mg holoenzyme/min for the homologous Synechococcus L8S8 and the hybrid Synechococcus L-pea S L8S8, respectively. The CO2 dependence for carbamylation of L8 was compared to that of L8S8 as a function of pH and CO2 concentration. The pH dependence indicated an apparent pKa for L8 of 8.28 and for L8S8 of 8.15, suggesting that S may influence the pKa of the lysine involved in carbamylation. The Kact for CO2 at pH 8.4 were similar for L8 (13.5 microM) and L8S8 (15.5 microM). L8 bound 2-[14C]carboxy-D-arabinitol 1,5-bisphosphate (CABP) tightly so that most of the bound [14C]CABP survived gel filtration. A major amount of the L8-[14C]CABP complex appeared as larger polymeric aggregates when eluted in the presence of E. coli protein.     

神经酰胺合酶-神经酰胺通路在青蒿琥酯抑制肝纤维化中的作用

目的: 探讨神经酰胺通路在青蒿琥酯(Art)抑制肝纤维化过程中的作用。方法: 将肝星状细胞(LX-2)分为细胞对照组、Art 350 μmol/L组、神经酰胺合酶阻断剂(FB1) 6 μmol/L组及FB1 6 μmol/L + Art 350 μmol/L合用组。每组7个复孔,作用24 h后,收集细胞及上清液进行检测。Western blot法测定神经酰胺合酶2(LASS2)、过氧化物酶体增殖物激活受体-γ(PPAR-γ)、Caspase-3蛋白的表达, HPLC-FLD法对神经酰胺(Cer)的含量进行测定,MTT法检测LX-2的增殖情况,酶消化法检测羟脯氨酸(Hyp)的含量。结果: 与细胞对照组比较,Art处理组神经酰胺合酶的蛋白表达、神经酰胺含量显著增加、LX-2细胞的增殖明显抑制、PPAR-γ和Caspase-3蛋白表达上调、羟脯氨酸分泌抑制 (P＜0.05);FB1处理组神经酰胺合酶的蛋白表达和神经酰胺含量显著降低、LX-2细胞增殖显著增加、PPAR-γ和Caspase-3蛋白表达下调、羟脯氨酸分泌增加(P＜0.05);与Art单用组比较,两药合用可显著降低Art对神经酰胺合酶蛋白的表达和神经酰胺含量升高的作用,减弱Art对LX-2细胞增殖、PPAR-γ、Caspase-3蛋白表达及羟脯氨酸分泌的作用(P＜0.05)。结论: 青蒿琥酯可通过神经酰胺合酶-神经酰胺通路增加细胞中神经酰胺水平,发挥抑制肝纤维化的作用。     

The discovery of 4-(3-pentylamino)-2,7-dimethyl-8-(2-methyl-4-methoxyphenyl)-pyrazolo-[1 ,5-a]-pyrimidine: a corticotropin-releasing factor (hCRF1) antagonist

Structure activity relationship studies led to the discovery of 4-(3-pentylamino)-2,7-dimethyl-8-(2-methyl-4-methoxyphenyl)-pyrazo lo-[1,5-a]-pyrimidine 11-31 (DMP904), whose pharmacological profile strongly supports the hypothesis that hCRF1 antagonists may be potent anxiolytic drugs. Compound 11-31 (hCRF1 Ki = 1.0+/-0.2 nM (n = 8)) was a potent antagonist of hCRF1-coupled adenylate cyclase activity in HEK293 cells (IC50= 10.0+/-0.01 nM versus 10 nM r/hCRF, n = 8); alpha-helical CRF(9-41) had weaker potency (IC50 = 286+/-63 nM, n = 3). Analogue 11-31 had good oral activity in the rat situational anxiety test; the minimum effective dose for 11-31 was 0.3 mg/kg (po). Maximal efficacy (approximately 57% reduction in latency time in the dark compartment) was observed at this dose. Chlordiazepoxide caused a 72% reduction in latency at 20 mg/kg (po). The literature compound 1 (CP154526-1, 30 mg/kg (po)) was inactive in this test. Compound 11-31 did not inhibit open-field locomotor activity at 10, 30, and 100 mg/kg (po) in rats. In beagle dogs, this compound (5 mg/kg, iv, po) afforded good plasma levels. The key iv pharmacokinetic parameters were t1/2, CL and Vd,ss values equal to 46.4+/-7.6 h. 0.49+/-0.08 L/kg/h and 23.0+/-4.2 L/kg, respectively. After oral dosing, the mean Cmax, Tmax t1/2 and bioavailability values were equal to 1260+/-290 nM, 0.75+/-0.25 h. 45.1+/-10.2 h and 33.1%, respectively. The overall rat behavioral profile of this compound suggests that it may be an anxiolytic drug with a low motor side effect liability.     

刺参养殖池塘颗粒悬浮物结构及其沉积作用

对不投饵刺参养殖池塘的颗粒悬浮物结构及其沉积量和沉积速度进行了研究.结果表明:(1)各池塘总颗粒悬浮物重量(TS)为(73.1±20.5)mg/L,变动于46.0～132.0mg/L之间;颗粒无机物(IS)含量为(50.6±12.6)mg/L,变动于28.5～76.5mg/L之间,占总颗粒悬浮物的69.13%;浮游生物干重(PZ)为(0.675±0.706)mg/L,变动于0.064～2.814mg/L之间,其中浮游植物干重(DWP)为(0.541±0.622)mg/L,变动于0.062～2.582mg/L之间,浮游动物干重(DWZ) 为(0.135±0.200)mg/L,变动于0.002～1.160mg/L之间;颗粒腐质为(21.91±19.59)mg/L,变动于4.18～83.86mg/L之间,占总颗粒悬浮物的比例为29.95%.(2)总颗粒物沉积量为(19.44±16.34)g·m-2·d-1,变动于6.33～91.98g·m-2·d-1之间,沉降速度为(0.30±0.30)m·d-1,变动于0.08～1.63m·d-1之间,新生态颗粒有机物沉积量为(5.09±7.30)g·m-2·d-1,变化于0.14～31.27g·m-2·d-1之间,总颗粒物沉积量和新生态颗粒物沉积量均是底层高于表层.     

Determination of cyclizine and norcyclizine in plasma and urine using gas—liquid chromatography with nitrogen selective detection

A rapid, sensitive and specific high-performance liquid chromatographic (HPLC) assay was developed for the determination of 8-chloro-6-(2-chlorophenyl)-4H-imidazo-[1,5-a]-[1,4]-benzodiazepine-3-carboxamide [I] and its 4-hydroxy metabolite, 8-chloro-6-(2-chlorophenyl)-4-hydroxy-4H-imidazo-[1,5-a][1,4]-benzodiazepine-3-carboxamide [II] in whole blood, plasma or urine. The assay for both compounds involves extraction into diethyl ether—methylene chloride (70:30) from blood, plasma, or urine buffered to pH 9.0. The overall recoveries of [I] and [II] are 92.0 ± 5.4% (S.D.) and 90.3 ± 4.9% (S.D.), respectively. The sensitivity limit of detection is 50 ng/ml of blood, plasma, or urine using a UV detector at 254 nm. The HPLC assay was used to monitor the blood concentration—time fall-off profiles, and urinary excretion profiles in the dog following single 1 mg/kg intravenous and 5 mg/kg oral doses, and following multiple oral doses of 100 mg/kg/day of compound [I].     

Mutations in the small subunit of ribulose-1,5-bisphosphate carboxylase/ oxygenase increase the formation of the misfire product xylulose-1,5-bisphosphate.

The small subunit (S) increases the catalytic efficiency of ribulose-1,5-bisphosphate carboxylase/oxygenase (EC 4.1.1.39) by stabilizing the active sites generated by four large subunit (L) dimers. This stabilization appears to be due to an influence of S on the reaction intermediate 2,3-enediol, which is formed after the abstraction of a proton from the substrate ribulose-1,5-bisphosphate. We tested the functional significance of residues that are conserved among most species in the carboxy-terminal part of S and analyzed their influence on the kinetic parameters of Synechococcus holoenzymes. The replacements in S (F92S, Q99G, and P108L) resulted in catalytic activities ranging from 95 to 43% of wild type. The specificity factors for the three mutant enzymes were little affected (90-96% of wild type), but Km(CO2) values increased 0.5- to 2-fold. Mutant enzymes with replacements Q99G and P108L showed increased mis-protonation, relative to carboxylation, of the 2,3-enediol intermediate, forming 2 to 3 times more xylulose-1,5-bisphosphate per ribulose-1,5-bisphosphate utilized than wild-type or F92S enzymes. The results suggest that specific alterations of the L/S interfaces and of the hydrophobic core of S are transmitted to the active site by long-range interactions. S interactions with L may restrict the flexibility of active-site residues in L.     

Preliminary studies on tissue culture and agrobacterium-medicated transformation of Brassica campestris ssp. chinensis]

The hypocotyls and cotyledons of the asepetic seedling of Brassica campestris ssp. chinensis L cv. Pudongaijiecai) were used as explants for tissue culture. Adventitious buds were differentiated on modified MS medium supplemented with TDZ 1-2 mg/L, NAA 0.2-1 mg/L and AgNO3 7.5 mg/L. The percentage of explants which formed buds of cotyledons was about 56%, and that of hypocotyls was about 37%. When the regenerated explants were transferred onto MS medium with 2 i.p. 5 mg/L and NAA 0.1 mg/L for two weeks, whole plantlets were obtained by culturing the regenerated shoots on 1/2 MS medium with NAA 0.1 mg/L. Agrobacterium tumefaciens strain (LBA 4404/PBI 121) carrying the GUS gene and Npt II gene was used for transformation. After 2 days of coculture, the hypocotyls and cotyledons were transferred onto regenerated medium containing CP 300 mg/L for bud formation. After 4-5 weeks, the differentiated buds were transferred onto selection medium with CP 200 mg/L and Km 10 mg/L for 1 month, then the green shoots were transferred onto the rooting medium containing Cef 100 mg/L and Km 20 mg/L. 4-5 weeks later, plantlets with Km resistance were obtained and some of them showed higher enzymatic activities of beta-glucuronidase than control ones.     

Regeneration of plantlets from mature embryo calli of Western Ghats land race cultivar of rice, Oryza sativa L

The Malnad region located in the Western Ghats of Karnataka is known for the cultivation of indigenous rain fed land race cultivar of rice. The present study was to investigate the callogenic and caulogenic potentialities of the two indigenous rice cultivar namely Karimundaga and Kanadatumba using dehusked mature embryo explants. For callus and shoot bud differentiation, the explants were cultured on Murashige and Skoog (MS) medium supplemented with 2,4-D (1-3 mg/L), IAA (1-2 mg/L), Kn (1-4 mg/L) and BAP (1-4 mg/L). The morphogenic potentialities of the two rice cultivar differed in texture of callus. In both the cultivar callogenic frequency was optimized at 1 mg/L 2,4-D concentration, it was 94% in Karimundaga and 58% in Kanadatumba. Supplementation of IAA either alone (1-2 mg/L) or in combination with Kn or BAP at 1 to 4 mg/L concentration of each induces shoot bud differentiation from the calli. In the cultivar Karimundaga caulogenic frequency was highest (10.60 +/- 2.55) at 1.0 mg/L IAA and 4.0 mg/L BAP concentration. While in the cultivar Kanadatumba highest number of shoot buds (7.90 +/- 2.69) was differentiated at 1.0 mg/L IAA and 4.0 mg/L Kn concentration. The calli derived regenerants were successfully acclimatized in the greenhouse and agro-morphological variations were evaluated. The growth characteristics and yield related parameters exhibited by in vitro plants were lower than the in vivo plants.