两个固氮相关的植物基因对烟草和水稻的转化及其表达

豆科植物凝集素基因和血红蛋白基因在对根瘤菌的识别作用和类菌体在低氧分压下的共生固氮中起重要作用。本文的目的是试图将这两个基因转移到非豆料植物烟草和水稻，使其能识别根瘤菌，探讨非豆科植物的共生和联合固氮的可能性。 构建了含有豌豆凝集素（P－Lec）基因、Parasponia andersonii血红蛋白基因、gus基因及植物选择标记潮霉素磷酸转移酶基因（hpt）的两个植物表达载体pCBHL和pCBHUL;同时，还构建了含有P－Lec基因、gus基因及植物选择标记PPT乙酰转移酶基因（bar）的植物表达载体pBBUL。在pCBHL中，CaMV35S启动子调控P－Lec基因的表达，而在pCBHUL和pBBUL中，该基因由玉米Ubiquitin 1启动子调控。 用农杆菌法将pCBHL导入烟草，得到53株再生植株，PCR检测表明转化频率为88％。用基因枪法分别将pCBHUL和pBBUL导入水稻幼胚或幼胚诱导的愈伤组织。转pCBHUL的材料共得到40株再生植株，经分子检测有18株分转基因植株，转化频率为0.9%。转pBBUL的幼胚愈伤组织经PPT筛选，只得到能分化出小芽的抗性愈伤组织。 PCR检测、Southern杂交表明P-Lec基因和Paraspoina血红蛋白基因都已经整合到转基因烟草及水稻的基因组中，转基因水稻植株中两个外源目的基因的拷贝数较高。Western杂交分析转基因植物中P-Lec基因的表达情况，结果表明该基因在转基因的烟草和水稻叶片中得到正确地表达。同时，GUS组织化学染色表明转基因烟草的嫩茎和幼根，转基因水稻的嫩叶和幼根中都有gus基因的表达。转基因烟草中外源基因的表达频率高于转基因水稻。T1代转基因烟草幼根的蛋白原位免疫杂交显示P-Lec正确定位于正在生长的幼根根毛的顶端，与对照豌豆中的P-Lec基因表达部位相一致。关于Parasponia血红蛋白基因，以前本实验室对转基因烟草和水稻的研究表明有转录水平的表达，国外实验室证实转基因烟草中有转译表达。 上述结果有可能为进一步研究转基因非豆科植物与根瘤菌的相互作用奠定一定的基础。

玉米的单倍体育种及基因转化的研究

玉米的单倍体育种，是利用花药培养或孤雌生殖产生单倍体后，进行人工或自然加倍，迅速获得稳定的新品种的育种方法。单倍体育种可以缩短育种时间，单倍体培养体系如果作为转基因的受体，可保证外源基因在后代中稳定遗传，而不发生分离。因此，玉米单倍体育种无论在实践中还是在理论研究方面都具有重大的意义。 本文针对玉米花药培养中长期以来未能解决的诱导频率低、基因型之间差异大、小苗移栽不容易成活等问题，重点探讨了各种因素对玉米花药培养的影响。结果表明：不同基因型之间的诱导频率差异明显，杂交种的诱导频率比纯系高，并选择出诱导频率高达20%的材料“中0198”;接种时花药中的花粉处于单核中期时，其诱导频率最高;采用液体培养基比采用固体培养基诱导频率提高一倍;培养基中加入0.5%的活性炭，可使诱导频率由5.25%提高到9.35%;15%的蔗糖浓度对玉米的花药培养是最适宜的，培养2周后，将培养基的蔗糖浓度从15%调整为10%，将明显提高诱导频率;培养基中高浓度的KT和低浓度的BA有利于诱导体细胞胚的发生，而低浓度的KT和高浓度的BA有利于诱导芽的发生;接种前将花药在4℃条件下进行低温预处理，可将诱导频率从3.13%提高到11.71%;培养基中添加2 mg/l的多效唑，可有效地促进小苗的生根;再生植株于冬季拿到海南种植，可明显提高移栽的成活率。 在玉米孤雌生殖的实验中，将未受粉的玉米雌穗接种在成份为N6 + 2,4-D 1mg/L + NAA 1mg/L + BA 1mg/L + CH 200mg/L + colchicine 2 mg/L + sucrose 5% + agar 0.7% 的培养基上。第一轮实验共接种了三个材料的26个雌穗（约3900个未受精的子房）。每种材料均有单性结实，诱导频率由高到低分别3.06%，2.29%，1.90%。直接获得了5株再生植株，通过染色体检查，发现其中3株为单倍体（n=10），另外2株为二倍体。移栽到土壤中后，有4株成活，其中一株二倍体植株能够正常开花、结实。得到的种子播种于实验田中，表现整齐一致，有纯系的特征，而且出现了2株白化苗。通过石蜡切片初步观察了孤雌生殖的胚胎发生过程，发现胚胎发生是从胚囊里的单倍体细胞起源的。第二、三轮实验又接种了10个基因型的玉米材料，证实了上述结果。 外源基因转导是利用生物技术进行玉米育种的一个有效途径。本文首次尝试了用离体子房注射法对玉米进行基因转化。首先构建了含有开花促进因子基因FPF1及植物选择标记抗除草剂基因pat的植物表达载体pFBR，采用离体注射培养法，取授粉24小时后的玉米雌穗，剥去苞叶，在超净工作台进行微量注射，然后切成小块接种在培养基上，在光照培养箱内培养，3-4周可直接获得种子或小植株。种子萌发后进行植株抗性筛选和分子检测，共注射了3个品种的47个雌穗（约16450个子房）得到再生植株109株，其中经PPT筛选有抗性的植株为23株，占再生植株的21.1%。经PCR检测，13株植株有阳性反应。但Southern杂交检测有杂带出现。出现杂带的原因、RNA水平的分子检测、转基因后代T1代的分子检测和早开花农艺性状的观察，由于时间关系没有完成，还需要进一步的实验。实验初步证明了离体子房注射法对玉米进行基因转化的可行性，而且与田间注射法相比，此方法具有省力省时，容易控制污染，转化效率提高的优点, 克服了玉米培养再生植株受基因型限制的困境, 将为玉米分子育种的基因工程提供更易行的手段。 同时，也为子房较大的其它植物的基因转化提供了方法。

BADH基因转化番茄提高耐盐性的研究

甜菜碱是植物在盐、干旱或其它胁迫下在细胞中迅速积累的一种相容性有机小分子化合物，它在细胞中的积累与植物抗盐性的提高密切相关。甜菜碱醛脱氢酶（BADH）催化甜菜碱醛转化为甜菜碱。我们将来源于耐盐植物山菠菜（Atriplx hortensis L.）的BADH基因通过农杆菌介导法导入‘百丽春’番茄(Lycopersicon esculentum L. ‘Bailichun’)中，并获得15株转化植株，PCR、Southern和Northern检测表明，其中的6株有外源BADH基因的整合，5株中BADH基因能够正常表达，但不同植株间BADH基因的表达水平和BADH酶活力有较大差异。对叶片电导率的测定表明，转基因植株比野生型的耐盐性有较大提高。T1代分析表明，检测的两个转基因株系后代遵循孟德尔分离规律，90mmol/L NaCl胁迫下种子发芽率提高了2~4倍，幼苗的苗高、根长和须根数三个指标均明显优于对照。部分T1代植株在水培条件下能够耐受180mmol/L NaCl胁迫。 植物耐盐的另一机理就是利用液泡膜上存在的转运蛋白将细胞内的有毒离子区域化。我们将已转入编码转运蛋白基因AtNHX1的番茄品种‘Moneymaker’（L. esculentum‘Moneymaker’）株系X1OEA1通过农杆菌介导法转入山菠菜BADH基因，以期获得转双基因耐盐番茄。目前已获得转基因植株，PCR结果证明部分抗性幼苗中已整合了BADH基因，其它各项分子检测正在进行中。

小麦TaRanBP基因功能的初步分析

小G蛋白（small GTPases）是真核生物中广泛存在的一类调节各种生命活动的信号分子。根据结构与功能的不同，小G蛋白家族成员可分成五个亚家族，分别为Ras，Rab，Rho，Arf和Ran。五类小G蛋白通过其活化态（GTP结合态）和非活化态（GDP结合态）的相互转换行使着各种功能。Ras GTPases在酵母和哺乳动物中调节细胞增殖过程; Rho GTPases调控肌动蛋白重组过程，并参与MAP 激酶的细胞信号转导过程等; Rab GTPases和Arf GTPases分别在膜转运过程中起着不同的重要作用;而Ran GTPases则在核孔位置调节着蛋白和RNA分子的运输过程。 小G蛋白附属蛋白调节着小G蛋白活化态与非活化态之间的转换，其中鸟核苷酸交换因子(guanine nucleotide exchange factors, GEFs)可以催化小G蛋白转换为GTP结合形式，即活化态;而GTPase 激活蛋白(GTPase-activating proteins, GAPs)和小G蛋白结合蛋白(small GTPases binding proteins)可以激活小G蛋白自身的水解活性，从而将其转变成非活化态形式。 相比其它小G蛋白，Ran GTPases及其附属蛋白在真核生物中的研究相对较少。已有的成果表明它们主要在核质运输过程中及对相应的信号转导途径起调节作用。而针对Ran GTPases及其附属蛋白在真核生物尤其是高等植物个体发育过程中的作用，目前报道还很少。 为了揭示Ran结合蛋白(Ran binding protein, RanBP)在植物发育过程中的作用，本文通过转基因手段对其功能进行 了研究。在此之前，本实验室已从小麦cDNA文库中成功克隆Ran结合蛋白基因：TaRanBP。该基因cDNA全长1035 bp，编码207个氨基酸。通过农杆菌介导叶圆片法，分别用正义、反义及TaRanBP与GFP融合蛋白等表达载体转化烟草，并成功获得转基因植株。亚细胞定位观察发现TaRanBP蛋白主要定位于细胞质内，尤其是在核膜附近富集。生理学和细胞学等方面的研究分析发现，TaRanBP基因在烟草个体发育过程中产生重要作用。过量表达TaRanBP基因的转基因植株在一定数量上表现出愈合的花冠筒上出现不同程度开裂，花冠筒上有附生舌状花瓣，及带有花瓣状颜 色的花萼等异常花表型。同时，转反义基因在一定程度上促进了转基因植株初生主根的生长（为对照烟草的2.3倍），而转正义基因烟草与对照烟草的初生主根长度差异不明显。用碘化丙锭(Propidium Iodide, PI)进行根部细胞染色。观察发现，不同的转基因烟草与对照烟草之间在根的各个不同形态区域的细胞大小差异不明显，推测根长的差异可能是由于整体细胞数目变化的原因导致。向重力性实验发现，转反义基因烟草幼苗较对照烟草的向重力性反应增加，而转正义基因的则表现为降低。激素吲哚乙酸(Indoleacetic Acid, IAA)的添加处理可以恢复转反义基因烟草的向重力性异常表型，而对转正义基因烟草几乎无影响。添加激动素(Kinetin, KT)的处理发现不同转基因烟草和对照烟草的向重力性均有减弱。观测后期，转正义基因的向重力敏感性较对照烟草得到恢复。测量不同转基因株系T1代幼苗鲜重，发现不同转基因烟草和对照烟草的幼苗鲜重动态变化在各个时间点有差异，且差异情况不尽相同。而不同转基因幼苗T1代幼苗可溶性蛋白含量较对照烟草有不同程度的下降。这种下降并没有影响转基因烟草的整体生长进程，开花期和结实情况与对照烟草相比也无明显变化。

SeNHX1和BADH基因提高植物耐盐性的研究

土壤的盐碱化问题已经严重影响到世界范围内许多重要作物的生产。培育耐盐作物是解决这一问题的最有效途经。利用耐盐相关基因的转化可以在不改变或很少改变植物其它性状的情况下提高植物的耐盐性，因此基因工程方法对于改良植物耐盐性及其机理的研究具有重要的意义。目前植物耐盐基因工程从调控渗透调节物质和盐离子区隔化两个方面开展了较多的研究。已经获得一些耐盐性提高的转基因植物。 本研究拟用耐盐性较强植物山菠菜中的甜菜碱合成关键基因BADH和盐生植物盐角草的液泡膜Na+/H+ anitiporter基因SeNHX1对模式植物烟草进行转化，以确定其各自在耐盐性方面所起的作用。同时，现有的研究表明植物的耐盐性是多基因控制的复杂性状，因此拟把SeNHX1和BADH 这两个涉及不同耐盐机理的基因构建到同一个植物表达载体上，以比较单基因转化和双基因转化在提高植物耐盐性方面的优劣。除此之外，并对已经转入BADH基因番茄的耐盐性和遗传稳定性分析进行了研究。 转BADH基因番茄已经稳定遗传到T4世代。通过对5个转BADH基因番茄株系在T0世代、T3世代和T4世代的分析，表明除了株系T4-3由T0世代的3个拷贝变为1个拷贝外，其余各株系拷贝数均没有发生变化。外源基因编码的酶活性和最终催化产物甜菜碱在盐分胁迫下都能较容易的检测到，说明外源基因在番茄基因组中的遗传是稳定的，没有发生丢失。在连续2个世代的耐盐性鉴定中，各转基因株系的耐盐性较为一致，均比野生型有了较大的提高。其中株系T4-5连续2年表现出了较低的减产率，株系T4-8也在连续的2年中表现出了最高的单株产量。盐分胁迫下转BADH基因各个株系比野生型有较高的K+和Ca2+含量，较低的Na+含量，转基因株系较野生型有较低的脐腐病果率。 通过SeNHX1、BADH单独转化以及构建双价载体共转化的方法获得了3种类型的转基因烟草。Southern和Northern 检测结果表明，外源基因已经整合到烟草基因组中，并得到了正确的表达。转BADH基因烟草在盐分胁迫下能检测到明显的BADH酶活性和甜菜碱含量。转基因烟草T0代对盐分胁迫、氧化胁迫的抗性均较野生型对照有较大的提高。转基因株系在200 mM NaCl胁迫下较野生型有较高的光合速度。百草枯处理过的野生型叶盘比转基因株系积累了更多的丙二醛，表明野生型受到了更大的氧化胁迫。 已经获得3种转不同基因烟草的T1代，且T1代具有较强的耐渗透胁迫能力。转基因烟草的T0种子均能在含100 mg/L 卡钠霉素培养基上发芽和正常生长，其中部分种子能够在含200 mM NaCl 培养基上发芽并能较好的生长，而野生型根本不能发芽。从200 mM甘露醇胁迫1周后，又转移到营养液中的生长1周的情况来看，转基因烟草能较快的恢复正常的生长，有新的叶子和根长出，而野生型却不能，同时转基因株系比野生型具有更大的单株鲜重。 转BADH基因番茄在遗传上是稳定的，并且其耐盐性有了较大的提高。双基因转化烟草的抗盐性要好于单基因转化，但SeNHX1基因转化要好于BADH基因转化。说明SeNHX1基因在提高植物耐盐性方面要比BADH基因有更强的功能，同时，也表明多基因转化在植物的耐盐改良方面可能是一个更为有效的方法。

内蒙古锡林河流域空间生态学研究

本文从物种和景观两个组织水平上研究了气候、土壤、地形等自然环境因子和人类活动因子对生物空间分布格局的影响。基于锡林河流域地理信息系统各环境因子的专题数据，利用空间异质性分析方法研究了锡林河流域环境因子的空间分布格局;基于锡林河流域野外调查数据，运用空间异质性分析方法研究了重要物种的空间分布格局，并采用典范对应分析(Canomc Correspondence Analvsis，CCA)方法分析了物种分布与环境因子的关系：基于锡林河流域地理信息系统各环境因子的专题数据，研究了锡林河流域植被斑块的空间格局特征及其与环境因子的关系，并采用典范对应分析方法分析了植被类型组成与环境因子的关系：基于内蒙古草原生态系统定位研究站放牧样地的样方调查数据．采用空间异质性分析方法，研究了放牧压力对物种空间分布格局的影响：基于多年的卫星遥感数据，采用建模和对比等方法，研究了定居放牧方式下植被状况空间变化规律及植被状况时空变化与人类活动、社会经济发展的关系。通过上述分析，得到的主要结论如下： 1、锡林河流域各个环境因子都具有自己的空间特征尺度，共同形成多尺度等级体系，按特征尺度的大小可以分为如下3个组： ·小尺度组(15km左右)：有机暖、全N的较小的特征尺度 ·中尺度组(30～50km)：T1，碳酸钙含量．PER、全N和海拔高度的较小的特征尺度 ·大尺度组(100km左右)：ANNR，PER、全N和海拔高度的较大的特征尺度多尺度等级的生态学意义是它反映生态变量异质斑块的镶嵌和包含特征，环境因子多尺度等级体系反映共性，具有普遍性：反映生态关系，具有生态学意义。 2、对物种空间异质性的Mantel检验和半方差分析得到了一致的结果产即羊草、糙隐子草和星毛萎菱菜在锡林河流域的空间分布呈现随机特征，而大针茅和冷蒿则表现为十分显著的格局特征。按分布格局的显著程度从大到小排列为冷蒿>大针茅>星毛萎菱菜>糙隐子草>羊草。理论半方差图显示大针茅和冷蒿的空间自相关域分别为30．447公里和30公里。物种空间分布格局是受自然条件、人类活动以及它们自身的生理生态特征综合决定的，物种自身的生理生态特征决定了它们对外界环境变化的适应性反应机制，而自然与人类活动这两种因素在空间的交错配置决定了物种适应性反应的方向和程度，从而综合导致物种空间分布格局的形成。 3、对锡林河流域物种分布与环境因子关系的CCA分析和交叉半方差方法分析显示：1)气候因子(11个指标)、土壤性状因子(3个指标)和地形因子(3个指标)对物种分布的贡献率分别为11．2％、9．5％和11％，三者总和为31．7％。2)各个环境因子对物种分布空间作用方向具有一致性，物种分布与环境因子几乎都在135。和157．5。两个方向上具有相对明显的相关性，从锡林河流域来看，这两个方向反映了气候、土壤以及地形从东南往西北的变化梯度方向。 4、对锡林河流域14个植被景观指数进行的PCA分析表明，锡林河流域植被斑块空间分布的物理特征主要表现在斑块的数目和大小方面，其次是在斑块的多样性方面，并可将它们分为4个组，分别反映锡林河流域植被斑块的不同特征： ·第一组：NP、PRD、LPI、MPS、PSSD和TE，主要反映景观斑块在数量和大小方面的特征; ·第二组：SHDI、SIDI、SHEI和SIEI，主要反映景观斑块的多样性特征; ·第三组：PSCV和[J]．主要反映景观斑块之间的相互邻接程度; ·第四组：MSI和AWMSI，主要反映景观斑块的形状特征。 MPS和PSSD两个指数与环境因子无论是在相关系数的性质还是显著程度上都保持了很好的一致性，它们与纬度(LAT)及可能蒸散率(PER)呈极显著的正相关关系，而与经度(LNG)、海拔高度(ALT)、年平均降水量(ANNR)及土壤有机质含量(0RG)呈极显著的负相关关系：平均形状指数(MSI)只与LAT呈显著的正相关关系;多样性指数和扩散毗连指数与任何一个环境因子都没有表现出显著的相关性。 5、锡林河流域植被分布与环境因子的关系CCA排序方法分析表明，气候因子(11个指标)、土壤性状因子(3个指标)和地形因子(3个指标)对植被分布的贡献率分别为19.8%、11.1%和14.5%，三者总和为45.4％。环境因子在植被和物种两个水平上的贡献率表现了相似的特点，自然环境因子不能完全解释植被的空间分布，人类活动的影响应该受到重视。 6、放牧压力对物种空间分布格局的研究表明： ·牧压对温带典型草原物种的空间分布格局有明显的影响。随着牧云的增大，属于原生群落物种的羊草与大针茅空间分布的随机性减小，空间自相关尺度逐渐增大;而对于退化过程中的入侵物种冷蒿和星毛萎菱菜，其空间分布的随机性逐渐增大．空间自相关尺度也呈增大趋势。在牧压胁迫超过一定水平时，冷蒿空间分布的自相关尺度开始下降，而星毛萎菱菜的空间分布格局则表现出强烈的随机性。 ·物种空间格局的变化是反映群落演替过程较为稳定的特征，适用于不同放牧条件下 群落之间的比较。 7、利用遥感数据对人类活动对植被影响的研究表明： ·定居放牧方式下，NDVI随定居点距离的变化格局经历了3个阶段。第一阶段，草场处于原生阶段，NDVI不随距离变化;第二阶段，定居点附近开始局部退化，NDVI随距离增加而增大：第三阶段，退化区域扩大，NDVI不随距离变化。 ·在草场局部退化阶段，NDVI随距离的变化呈对数函数规律，定居点的放牧区具有放牧半径、原生NDVI值、NDVI变化率等特征。根据这些特征、NDVI对数规律以及NDVI与地上生物量的关系可以推测定居点的总载畜量。 ·锡林河流域从87年到85年NDVI值降低最大的区域为流域的中部和南部，这与这一区域人类活动强度以及社会经济发展具有密切关系。

Effect of stocking density on growth of tiger shrimp (Penaeus monodon Fab.) fed on commercial formulated diets

Three different stocking rates in a semi-intensive pilot shrimp project was adopted in duplicates of three treatments designated as T1, T2 and T3 having initial per meter square stocking density of shrimp of 40, 44 and 51 respectively of 0.025g size post larva. The study was conducted for 84 days. Commercial pelleted diets designated as starter - 1, 2, 3 and grower were fed at a satiation level during the study period with a feeding frequency of 4 to 5 times per day. Feed rationing was based on the survivability, body weight and tray checking. 5-25% of the pond water was exchanged daily. Sampling was done for growth after every 2nd week. Monthly sampling was done for mortality in the ponds. Mean weight gain of the shrimp in treatments T1, T2 and T3 were 16.96 ± 1.14, 16.04 ± 1.38 and 14.08 ± 1.17g respectively and T1 with a low stocking density showed a significantly best growth among the treatments. Total mortality in treatments T1, T2 and T3 were as 30.00, 39.77 and 31.37% respectively. Significantly higher feed conversion ratio (FCR) of 1.87 was obtained with shrimp in treatment T3 followed by shrimp in T1 and T2 with FCR values of 1.70 and 1.41 respectively. A positive correlation of growth and salinity was observed during the study. Total production per unit area was the highest in the treatment T3 (4928 kg/ha) and followed by T1 (4747 kg/ha) and T2 (4251 kg/ha). The result show significantly negative correlation between individual growth and density.

Culture feasibility of african catfish (Clarias gariepinus Lin.) fry in glass tank and synthetic hapa system using supplemental diets

An experiment was carried out with 1 0 days old Clarias gariepinus fry over a period of 42 days to determine the effects of different feeds on growth and survival of African catfish fry in glass tanks. The experiment was designed into four treatments each having three replications. Thus treatment 1 (T1) was named as Tank Tubifex (TT) and treatment 2 (T2) as Tank Sabinco (TS), treatment 3 as Pond Tubifex (PT), and treatment 4 (T4) as Pond Sabinco (PS). Live Tubifex (protein levels 64.48%) was supplied to treatments 1 and 3 and rest of the treatments were supplied Sabinco starter-1 (protein levels 40.13%). The highest and the lowest growth in total length and weight were 12.90cm, 18.77g and 6.17cm, 4.04g recorded from the treatments 3 and 2, respectively. Growth of catfish fry under treatment 3 in terms of both length and weight were significantly higher (P<0.01) than those of the other treatments. However treatment 2 showed the significantly lowest (P<0.01) growth performance among the various treatments. The highest survival rate (92%) was also obtained with treatment 3. Tubifex proved to be the best larval feed in respect of growth and survival rate.

Effects of culture treatments on HUFA levels in Artemia cysts

Artemia cysts (of GSL, Utah, USA origin) were produced from the modified traditional solar salt works of Bangladesh during winter months through different feeding/fertilization treatments (T1, T2, T3, T4 ) were analyzed to understand the effects of treatments on their fatty acid profile. Palmitic, Linolenic, Eicosapantaenonic and Docohexaenoic acids (mg/g. DW) were found highest for the cysts in T1 (16.0% ±1 .36), T2 (14.7% ±0.47), T3 (4.7% ±0.40) and T4 (0.7% ±0.06) treatments, respectively. High amount of 18:3(n-3) acids in the cysts of all sources proves to be freshwater type of the cysts. The presence of marine type essential fatty acids in the cysts of all sources were found low for 20:5n-3 (3.7-4.7%) and very low for 22:6n-3 (0.09-0.7%). No significant variation was observed for 16:0 acids within the treatments, but for 18:3(n-3) acid, the variation was found highly significant (P= 0.0052) between T2 and T4 treatments. For 20:5(n-3), only variation between T2 and T4 was found insignificant (P=0.1161), but between other treatments, significant variation was observed between T2 and T4 (P=0.0241), T2 and T4 (P=0.0022) and T1 andT4 (P=0.0161). No significant variation was found in other treatments.

Suitability of rich-fish culture under mono and polyculture systems in the boro rice ecosystem

An experiment was conducted in farmers’ fields under Paikgacha thana, Khulna to study the suitability of integrated rice-cum-fish culture. Three treatments namely T1 (Puntius gonionotus), T2 (Puntius gonionotus and Cyprinus carpio) and T3 ( Cyprinus carpio) were included for the study each having three replicates. The fish were stocked at a density of 3750/ha in all the rice plots. The physicochemical parameters of water viz., water depth, temperature, dissolved oxygen, pH, salinity, nitrate and phosphate etc. recorded during the study period were found within optimum range. Of the two cultured species C. carpio attained the highest average individual weight (160g) and survival (81.06%). With respect to biomass and income, highest average fish production and net profit per hectare (306.74kg and Tk. 8177.91) were obtained in T2 and the lowest (184.17kg and Tk. 2049.41) obtained in T1 and a significant variation (p<0.05) in fish production was observed among the treatments while for rice production, it was insignificant. The cost benefit ratio of fish production found were 1:1.29, 1:2.14 and 1:1.90 for T1, T2 and T3, respectively.

Effects of cowdung application on the production of mud crab (Scylla serrata Forskal) in brackishwater pond

A culture experiment of mud crab for 84 days was conducted in earthen pond at Brackishwater Station, Bangladesh Fisheries Research Institute, Paikgacha, Khulna. The aim of the experiment was to study the effects of cowdung application on crab production and water quality. There were three treatments as without cowdung (T1), 500 kg cowdung/ha/fortnight (T2) and 750 kg cowdung/ha/fortnight (T3) with three replicates for each. The result was evaluated on the basis of growth, production, survival rate etc. The experimental months were mid-April '96 to mid-July '96. To maintain a good water quality, water was exchanged in every spring tide. The range of salinity during the experiment was 8-19 ppt. Trash fish and fresh shrimp head were used as feed on raw basis in every alternate week at a rate of 8% body weight of crab at the same time rice bran and wheat flour were used at a rate of 2% body weight as a source of carbohydrate. The production of T1, T2 and T3, were 720.35 kg, 862.16 kg and 669.19 kg/ha respectively. Though the effects of cowdung on production of crab is insignificant but in terms of production, survival rate and growth, the study suggest that the application of cowdung in addition to feed can be recommended for mud crab culture at a rate of 500 kg/ha/fortnight.

Predation by Channa striatus (Bloch) on Rana tigrina (Daudin), Puntius gonionotus (Bleekeer) and Labeo rohita (Hamilton) in the laboratory

The growth performance of a predatory snakehead, Channa striatus was tested by supplying tadpoles of Rana tigrina and fingerlings of Puntius gonionotus and Labeo rohita as prey for a period of 21 days in aquaria. Prey consumption by C. striatus was significantly different (P<0.05) for different prey used (T1 - R. tigrina, T2 - P. gonionotus, T3 - L. rohita). Tadpoles of R. tigrina were preferred by the predator (C. striatus) over P. gonionotus and L. rohita although tadpole is nutritionally inferior to each of P. gonionotus and L. rohita. Each predator rayed on 50-330 mg per day per g of their body weight. Fish preyed on tadpoles also showed the highest growth. Significant difference in weight gain was found between T1 and T2 and also between T1 and T3 but no difference was found between T2 and T3. Food conversion ratio (FCR) was found to be lowest in treatment T3 followed by the treatments T2 and T1 respectively.

Effect of submerged aquatic vegetation on growth and survival of Penaeus monodon (Fab.)

An experiment was conducted in brackish water ponds with transplanted aquatic weed (Najas sp.) to study the effect of submerged aquatic vegetation on culture of Penaeus monodon. Out of six ponds three were without any aquatic vegetation (T1) while in other three ponds weeds were planted covering 40% of the pond bottom (T2). Hatchery produced post larva of P. monodon (0.006 g) were stocked in all ponds at a density of 40,000/ha. Shrimps were fed twice daily with commercial and formulated feed prepared from locally available ingredients. After 105 days of rearing shrimp of highest average weight (63.18g) was obtained from T2 with a survival rate of 25.90% and the total production was obtained at 654.54 kg/ha. The average weight, survival and total production of shrimp in T1 were 35.0 g, 28.76% and 405.63 kg/ha, respectively.

Effect of stocking density on the growth of Thai pangas, Pangasius sutchi (Fowler) in net cage fed on formulated diet

A three month long experiment was conducted to observe the effect of stocking density on the growth of Pangasius sutchi in net cages. The size of each cage was 1m³.The three stocking densities used were 40, 50 and 60 fishes/m³ and designated as treatment T1, T2 and T3 respectively. Each treatment had three replicates. All the fishes were of same age group having mean length and weight of 7.13 ± 1.37 cm and 2.46 ± 0.12 g respectively. The fish in all the net cages were fed a diet containing 34% protein. The result of the study showed that fish in the treatment T1 stocked at the rate of 40 fish/m³ resulted the best individual weight gain followed by T2 and T3 respectively. The specific growth rate (SGR) ranged between 3.51 and 3.09, the food conversion ratio (FCR) values ranged between 1.73 and 2.04 with treatment T1 resulting the lowest FCR. The protein efficiency ratios (PER) values were 1.69, 1.16 and 1.43 for treatment T1, T2 and T3 respectively. There was no significant (P>0.05) variation among the survival rates of fish which ranged between 92 and 95%. The net productions in different treatments were 2189, 2343, and 2283g for treatment T1, T2 and T3 respectively. The result of the present study indicated that the best individual growth of P. sutchi was obtained at a density of 40 fish/m³ but the highest total production was obtained at a stocking density of 50 fish/m³ in net cages.

Minimization of cannibalism of African catfish (Clarias gariepinus Burchell) larvae in indoor culture system

For minimizing cannibalism of African catfish (Clarias gariepinus) larvae two trials for a period of 14 and 15 days respectively in four aquaria of size 120x49x32 cm³ were conducted. Seven days old African catfish larvae with an initial total length and weight of 7.84 (±0.40) mm and 4.40 (±1.18) mg respectively in the first trial and similarly 7.52(±0.61) mm and 3.98 (±0.56) mg in the second trial at the rate of same stocking densities of 2500 larvae in each aquarium were stocked in both trials. Cannibalistic larvae were separated by using grader frame from each treatment at 7 days and 5 days interval during first and second trial respectively. Two mesh sizes i.e., 5 mm and 7 mm were used in the grader frame in both trials. Survival rate was significantly higher in T1 than that of T2 in each trial. Grading of larvae with 5 days interval resulted higher survival rate than that of 7 days interval.