分子运动性预测顽拗性种子麻栎离体胚轴适宜贮藏条件初探

顽拗性种子在分类上分布广泛。由于顽拗性种子贮藏特性极为特殊，对脱水和低温均十分敏感，易受损伤，所以种子寿命很短，因此在植物种质资源保存领域，顽拗性种子的保存一直被列为重点研究课题，至今仍难以找到顽拗性种子的适宜贮藏条件，或该类种子保存通行的方法，仅推测长期保存顽拗性种子种质资源最有前途的方式是超低温保存，顽拗性种子资源保存是一个世界性难题。 本文试图使用分子运动性预测顽拗性种子离体胚轴的适宜贮藏条件。选用壳斗科植物麻栎种子为实验材料。应用电子顺磁共振波谱仪和自旋标记技术，以硝基氧探针CP为标记物，检测到室温下麻栎种子离体胚轴脱水过程中分子运动性的变化。含水量0.7 g H2O/g DW至0.64 g H2O/g DW范围是细胞质粘度的转折区域，低于这个含水量区域，细胞质粘度骤然上升，推测这个区域是室温下保存离体胚轴的适宜含水量下限。 通过变温电子顺磁测定，找到离体胚轴含水量在0.43 g H2O/g DW至1.02 g H2O/g DW范围内，分子运动性的临界温度和玻璃态相变温度所在区间。根据分子运动性随温度变化规律，预测含水量为0.69 g H2O/g DW的麻栎种子离体胚轴适宜贮藏温度约为-50 ℃。 根据EPR实验结果，将胚轴脱水，并根据实验条件选择不同温度进行低温贮藏验证实验，应用颜色观测法和TTC存活力鉴定法，初步验证分子运动性理论可以应用在顽拗性种子离体胚轴的低温贮藏研究中。 本研究说明顽拗性种子资源的保存有可能通过离体胚轴的低温贮藏实现，用分子运动性预测低温贮藏的适宜条件，很可能为上述问题的解决提供了有效的技术参数。

利用DSC和EPR技术研究杜仲种子的贮藏条件

种子贮藏稳定性对于种质资源的长期保存具有重要意义，目前关于种子贮藏的最新理论为玻璃态理论，该理论认为种子的玻璃化有利于种子的长期贮藏。当种子处于玻璃态时，玻璃化物质的高度粘滞性降低了种子细胞内分子流动性，阻止了细胞质中分子的扩散，从而减少老化过程中细胞结构的损伤和化学组分的变化，延缓种子老化劣变反应速率，延长贮藏寿命。评价玻璃态的一个重要指标是玻璃化转变温度，当种子贮藏于玻璃化温度或以下10℃～30℃范围内时，种子具有最佳的贮藏稳定性。因此，检测种子的玻璃化转变温度对于种子的长期有效贮藏具有重要指导意义。 本研究将差示量热扫描技术（DSC）与电子顺磁共振波谱仪技术（EPR）应用于杜仲种子玻璃化转变温度方面的研究。在DSC方法中，选用4.4%～31.6%含水量范围的杜仲种胚分别进行了DSC图谱扫描。EPR方法选用3-羧基-2,2,5,5-四甲基吡咯烷-1-氧（3-carboxy-2,2,5,5-tetramethylpyrrolidine-1-oxyl，CP）和2,2,6,6-四甲基哌啶（4-hydroxy-2,2,6,6-tetramethyl-1-piperidinyloxy，TEMPO）作为探针标记杜仲种胚， 利用EPR技术测定不同含水量杜仲种胚的分子运动，通过对EPR图谱参数的分析计算，最终确定不同含水量杜仲种胚的玻璃化转变温度。 DSC实验结果显示，含水量为22.3%、28.0%、31.6%的杜仲种胚在0℃ 左右出现了一个水的熔融峰。该熔融峰的面积代表了自由水含量的多少，随着种胚含水量的降低该熔融峰面积减小。4.4%～31.6%含水量范围的杜仲种胚在-28℃左右还出现了一个熔融峰，推测此峰为杜仲种胚中某类物质熔融所形成的熔融峰。然而在此曲线上我们未观察到标志玻璃化转变的“台阶”出现。 CP-EPR实验的结果表明，利用EPR测定得到含水量为4.4%～11.6%的杜仲种胚在-110℃～20℃温度范围内，同一含水量的杜仲种胚随着温度的升高，分子运动速率加快;在同一温度条件下，高含水量的种胚比低含水量种胚的分子运动速率快。通过CP-EPR波谱两外缘峰最大距离（2Azz）的测定和数据统计分析，得到含水量为4.4%、5.7%、8.6%、10.3%、11.6%杜仲种胚的玻璃化转变温度分别约为44℃、25℃、4℃、-31℃、-43℃。可以把测定的杜仲种胚的这几个含水量的玻璃化转变温度与杜仲种子贮藏相结合，用于指导杜仲种子的贮藏。 TEMPO-EPR实验测定分析得到含水量为2.1%、3.4%、4.8%、8.3%、11.2% 的杜仲种胚的玻璃化转变温度分别为-21℃、-18℃、-24℃、-20℃、-27℃，玻璃化转变温度随含水量升高其变化的规律不明显，这与CP-EPR实验测得的结果有着较明显的差别。通过分析，认为对于脂质含量较高的杜仲种胚，随着含水量的降低，作为标记化合物的TEMPO随着脱水进入脂相，从而不能真实反映出不同含水量种胚的分子运动情况。与TEMPO标记相比，CP标记可能能够更真实地反映不同含水量杜仲种胚细胞质分子运动的情况，根据其分子运动情况得到的玻璃化转变温度更准确。

高等植物PSI ,PSII反应中心及天线系统色素蛋白复合体结构与功能的研究

一、实验证明了Cd H、Mg“在小麦类囊体膜上色素蛋白复合体的解聚和再聚合过程中，具有不同的作用。因此，二阶阳离子对激发能在光系统间的分配调节作用，可能不能仅仅用“静电现象”(Barber(1980))去解释。分析表明，在Ca2+作用下与PSII内周天线CP-47，GP-43多肽结合的L H C II和LHClb是来自间质膜区的PS I系统的。从PSI迁移到P S II的捕光色素蛋白，增加了PSII的捕光截面，从而促进了激发能有利于PsII分配。 二、Ca2*、Mg2+对小麦和菠菜类囊体膜光谱性质的影响有所差异。Ca2+对小麦类囊体膜光谱性质的影响还可以随着介质中Ca2+的消除而消除。同小麦类囊体膜相比，菠菜PSII以及LHCII更为集中在基粒区域，这可能是菠菜类囊体膜强Fv以及高F888／F735，F89H／F735比值的原因。因此，Ca2+，HgH对激发能在光系统间分配的调节作用是依赖于光系统间激发能及天线色素蛋白的分配状况的。 三、对菠菜叶中分离的PSII-RC: D1-D2-cyt b55g复合物进行的低温荧光发射光谱的研究表明，这一复合物可能具有F681和F684两种波长的低温荧光发射，但它们通常并不是同时存在，而是取决于Ca-670与Ca-680 Chla分子的相对含量的。PSII-RC内周无线GP-47，GP-43多肽的存在是D1-D2-cyt b559复合物低温荧光发射红移的原因;而D1一D2cyt b559复合物的不稳定性则与其低温荧光发射的蓝移现象有关。 从蕹菜叶中分离的Dl—D2-cyt b559复合物的F 381低温荧光发射也是由其相对含量较高的C．i-6 7 0 Chla分子的存在决定的。对蕹菜D 1一D 2-cyt b559复合物中的分析还表明，F 681的低温荧光发射直接来源于Di／D2复合物，而415nm处相对较强的吸收，则可能主要是与Pheo的存在有关的。 四、多肽分析与光谱分析的对照表明，CP-26内周天线多肽可能是PSII中F695低温荧光发射的真正来源。 五、实验分析了蔗糖密度离心分离的LHClI和PSI颗粒。结果排除了CP-27多肽（以及CP，一2 5，GP-47，CP -4 3多肽）具有F695低温荧光发射的可能，因此支持了CP-26多肽是PSII中F695低荧光发射来源的看法。对PsI颗粒的分析表明，P700的存在可能是与PSI-RC中较大的Sub-I亚基相联系的。 六、根据以上的研究结果，提出了PSI，PSII在类囊体膜上的结构模式，并对其内容进行了分析和讨论。

水分胁迫对光合作用影响的研究

1．水分胁迫降低了水稻叶片的光合速率。轻度及中度水分胁迫下，气孔的限制是光合速率下降的主要原因;而严重水分胁迫下，叶肉细胞光合能力的降低是光合速率下降的主要原因。叶肉细胞光 合能力的降低可能是由于光合量子效率、光合电子传递速率和光合磷酸化活力及羧化效率下降所致。 2．水分胁迫降低了小麦叶片的可变荧光产量、可变荧光淬灭速率、荧光上升互补面积，叶绿体的室温荧光产量、可变荧光产量以及DCMU作用下的小麦叶片及叶绿体的可变荧光产量，表明光系统Ⅱ受到了伤害。光系统Ⅱ氧化侧的人工电子供体(DPC) 能部分恢复受抑制的叶绿体可变荧光和光系统II的电子传递速率，说明水分胁迫对光系统Ⅱ的损伤不仅位于氧化侧，也可能在反应中心上。 3．水分胁迫抑制了激发能向PSII的传递;降低了Mg2+对叶绿体可变荧光及激发能在两个光系统间分配的调节能力。水分胁迫使PS II内外周天线色素蛋白复合体（CPa和LHCⅡ）和PSⅠ叶绿素a蛋白复合体(CPⅠ、CPⅠa和CPⅠb)含量下降，其中以LHCⅡ降低幅度最大。从类囊体膜多肽分析结果，发现25KD多肽随着水分胁迫的加剧其含量显著降低。 4．水分供应充分及轻度和中度水分胁迫下，高氮素营养对水稻光合作用有明显的促进作用;而严重水分胁迫则削弱了高氮营养对光合作用的促进作用，使高氮营养水稻叶片在严重水分胁迫下的光合速率反而比低氮营养叶片低，这可能与严重水分胁迫下，高氮叶片水势、气孔导度、光合羧化效率、光合量子产量及RuBP再生速率比低氮叶片有更大的下降有关。两种氮素营养水平下，轻度和中度水分胁迫均提高了叶片的水分利用效率;而严重水分胁迫则使叶片的水分利用效率降低，但始终是高氨营养水稻叶片的水分利用效率高于低氮叶片。

马铃薯Y病毒外壳蛋白基因的导入以及转基因马铃薯植株的大田攻毒

应用四种不同的马铃薯试管微型薯诱导体系生产试管微型薯，通过比较建立了—种有效的试管微型薯诱导系统。这种诱导系统使用液体培养基，具有试管微型薯发生频率高、薯块体积大和微型薯形成早的特点。同时，此系统使用的培养基成分成本低、方法简便以及所用设备简单，适用于容器内大批量生产马铃薯试管微型薯以及马铃薯种质资源的保存。 对银离子在马铃薯叶片组织培养过程中对愈伤组织诱导或芽分化和再生的影响作了研究。结果表明银离子通过抑制叶片组织培养过程中形成的乙烯与其受体的结合从而促进芽再生，但是其对叶片愈伤组织的诱导无显著效果。银离子的这些作用在通过同时应用2，4-D而明显表现出来。2，4-D通过促进乙烯的生物合成而降低银离子的促进作用，两者则通过对乙烯的调节而影响马铃薯叶片的愈伤组织诱导和芽分化再生。 将马铃薯Y病毒外壳蛋白基因通过根癌农杆菌双元载体系统导入马铃薯品种Desiree、K4和Favorita，获得了若干转基因株系。除了K4品种中—转化株系具有非正常生长形态外，其余转基因植株都生长发育正常。由此表明以根癌农杆菌介导的马铃薯转化中，构建于双元载体上的外源目的基因是随机进入并整合到受体细胞的染色体上。具有畸形生长性状的转基因植株的产生说明了PVY CP基因的整合可能干扰了控制正常生长发育、尤其是形态建成的基因表达。 在转化试验中，应用了试管微型薯薄片、茎切段和叶片三种外植体作为转化材料。对转化过程中农杆菌对外植体的侵染时间、共培养时间、外植体的类型以基因型对转化频率的影响作了比较研究。发现以试管微型薯薄片和茎切段作为受体的最佳侵染时间是十分钟，而叶片则为五分钟，三种外植体的最佳共培养时间皆为四天。在各种处理的最佳条件下，Desiree比K4具有相对较高的转化频率，表明马铃薯Desiree比K4在转化反应上更温和或顺从。 通过比较几种由不同统计得出的农杆菌介导的转化频率，认为使用“净转换频率”(Net Transformation Frequency)能更精确地表达马铃薯的转化效率。而在以前的报导中还没有—种统一的、并且能被广泛接受和使用的表达转化效率的参数或指标。． 以叶片作为起始材料的转化具有较高的转化频率。在转化外植体的植株再生过程中应用了2，4-D和AgN03两种乙烯调节剂分别于愈伤组织诱导和芽分化再生阶段，使其产生高频的植株再生。尤其是它的净化频率明显高于其它外植体的转化频率，并且无显著品种之间的差异，具有高效马铃薯转化系统的特征。 以聚合酶链式反应(PCR)检测转化再生植株得到的结果与DNA杂交(Southern blot analysis)的鉴定结果比较，结论是相同的。由此表明在以农杆菌介导的马铃薯转化试验中，PCR可被用于证实外源目的DNA的导入，它以简便、迅速的特点帮助节省时间以及提供及时的转化证据。 对三个马铃薯品种的一系列转基因株系在大田条件下进行了攻毒试验．最后从Favorita 品种中筛选出了两个抗性较强的无性系，它们具有明显较低的病毒侵染发生频率以及正常的生长发育性状，具有很大潜力成为生产上推广应用的抗病新品种。

Effect of feeding three different formulated feeds having different protein levels on the growth of angel fish (Pterophyllum scalare) juveniles

Experiment was conducted to study the effect of dietary protein level on growth and nutrient utilization by angel fish (Pterophyllum scalare) juveniles. Fifty-four juveniles (average wt. 2-2.5 g) were equally divided in three treatments with each of three replicates. Three formulated diets with graded protein levels, T1 (35% CP), T2 (40% CP) and T3 (45% CP) were fed to juveniles for 45 days. A trend of higher weight gain %, SGR, FER and PER was found with the increased CP level in the feed. Feed intake was similar in all the groups. T3 group fed with 45% CP registered highest weight gain % (43.26 ± 2.07), SGR (0.78 ± 0.04), FER (0.29 ± 0.01), which were significantly higher (P<0.05) than the T1 and T2 groups. Protein digestibility of T2 and T3 groups was significantly higher than the T1 group. Survival was similar in all the experimental groups. Diet with 45% CP with protein energy ratio of 112.62 mg protein/K cal. is ideal for juvenile angel fish for indoor rearing.

Observation on some of the environmental parameters and feed quality of selected golda (Macrobrachium rosenbergii) farms in Bangladesh

A study was conducted to ascertain the existing farm water, effluent and feed quality of selected giant freshwater prawn farms from major prawn farming areas (Bagerhat Sadar, Noakhali Sadar and Mymensingh) of Bangladesh during July to November 2005. Water quality parameters such as the mean values of dissolved oxygen, alkalinity, nitrite-nitrogen, phosphate-phosphorus and ammoniacal nitrogen did not show any significantly differences among the farming areas. Whereas significant differences (p < 0.05) were observed in the mean values of temperature, secchi disc visibility, pH and chlorophyll a. However, all the water quality parameters in the farming areas were within the suitable range for prawn culture. There was no significant variation in nutrients concentration of discharged effluent among the prawn farming areas. All of the nutrients measured in effluent water were within the acceptable range and did not seem to pose a direct threat to the recipient environment. The analysed crude protein contents of commercial CP, Quality and Saudi-Bangla prawn feeds were 31.84%, 27.21% and 28.97%, respectively, whereas all analysed farm made feeds were less than 25%. The other nutrients of prawn feeds varied largely with the source of feeds and ingredients used to prepare feed. The annual yield of prawn varied from 320.4 to 512.6 kg/ha (mean 412.3 kg/ha) depending on the management system.

Effects of iso-nitrogenous and iso-phosphorus fertilizers as nutrient sources on carp polyculture in Bangladesh

A 120 day long experiment was conducted to find out the effects of cow manure with urea and triple super phosphate (CUT), poultry manure with urea and triple super phosphate (PUT) and cow manure with poultry manure (CP) having similar quantities of nitrogen and phosphorus on pond productivity and fish yield. The stocking fish were rohu (Labeo rohita), catla ( Catla catla) and mrigal ( Cirrhinus mrigala) in each treatment pond at the rate of 10000/ha. All ponds were fertilized fortnightly at the rate of 4000 kg/ha cow manure with 62 kg/ha urea and 65 kg/ha TSP, 2700 kg/ha poultry manure with 62 kg/ha urea and 16 kg/ha TSP, and 4000kg/ha cow manure with 2700 kg/ha poultry manure for the treatment CUT, PUT and CP respectively. Each treatment contained an iso-nitrogen and iso-phosphorus of 56 kg and 46 kg respectively. Though the physico-chemical parameters were more or less similar in all ponds, the chlorophyll-a content and abundance of total plankton were significantly higher (P< 0.05) in the ponds receiving the fertilizer treatment of PUT than those of other treatments. Final growth as well as per unit production of fish of treatment PUT (1773 kg/ha) was significantly higher (P< 0.05) than that of treatment CP (1528 kg/ha) followed by that of treatment CUT (1336 kg/ha). The over all results showed that poultry manure with urea and triple super phosphate proved to be superior to cow manure with urea and triple super phosphate, and poultry manure with cow manure, even when nitrogen and phosphorus content was similar, in carp polyculture system under prevailing conditions.

Effect of feed quality restriction followed by realimentation on growth, nutrient utilization, biochemical changes and haematological profiles of Indian major carp, rohu (Labeo rohita H.)

To investigate the effect of protein restriction with subsequent re-alimentation on nutrient utilization, hematological and biochemical changes of Indian major carp, Rohu (Labeo rohita H.), 150 acclimatized Rohu fingerlings (average 20.74 ± 0.13 g) divided into five experimental groups (30 fingerlings in each groups with three replications with 10 fingerlings in each) for experimental trial of 90 days using completely randomized design. Control group (T sub(CPR)) was fed with feed having 30% crude protein at 3% of body weight for 90 days trial period. Other experimental groups T sub(1PR) was alternatively 3 days fed with feed having 20% CP and 30% CP at 3% of body weight, T sub(2PR) was alternatively 7 days fed with feed having 20% CP and 30% CP at 3% of body weight, T sub(3PR) was alternatively 15 days fed with feed having 20% CP and 30% CP at 3% of body weight and T sub(4PR) was alternatively 25 days fed with feed having 20% CP and 30% CP at 3% of body weight during 90 days trial period with daily ration in two equal halves at morning and afternoon. It was noticed that retention of different nutrients was almost similar among all treatment groups indicated improvement of digestibility of nutrients might not be the mechanisms for recovery growth in carps. Increased percent feed intake of body weight (hyperphagia) (4.14 ± 0.30 or 4.94 ± 0.46 and 3.33 ± 0.29), improved specific growth rate (1.86 ± 0.09 or 2.26 ± 0.05 and 1.43 ± 0.01), absolute growth rate (1.57 ± 0.08 or 1.84 ± 0.18 and 1.36 ± 0.12), protein efficiency ratio (1.19 ± 0.11 or1.16 ± 0.12 and 1.05 ± 0.09) were the important mechanism showing better performance index (21.60 ± 1.09 or 23.80 ± 0.21 and 19.45 ± 0.37) through which the experimental groups which were protein restricted and re-alimented at 3 or 7 days alternatively during 90 days trial period could able to compensate the growth retardation and to catch up the final body weight of control (128.68 ± 11.53 g/f) but other experimental groups failed to compensate during 90 days trial period. Result of the present study indicated that deprived fish i.e., fish received alternate 3 or 7 days protein restriction and re-alimentation showed recovery growth had still lower values of Hb (10.21 ± 0.02, and 9.88 ± 0.04 g/dl), hematocrit value (30.62 ± 0.05 and 26.64 ± 0.11%), total erythrocytic count (3.40 ± 0.01 and 3.29 ± 0.01 X10super(6) mm³), plasma glucose (126.93 ± 0.20 and 126.67 ± 0.05 mg/dl), total plasma lipid (1.04 ± 0.01 and 1.02 ± 0.01 g/dl) and liver glycogen (290.10 ± 0.80 and 288.99 ± 0.95 mg/kg) in comparison to control (10.56 ± 0.08 g/dl, 31.68 ± 0.24%, 3.52 ± 0.03 X10super(6) mm³, 128.23 ± 0.25 mg/dl, 1.07 ± 0.01g/dl and 292.00 ± 0.23 mg/kg) at the end of 90 days trial but total plasma protein in deprived group was compensated with advancement of trial period. All hematological and biochemical parameters studied were proportionately lowered in the experimental group got higher degree of deprivation. These findings suggested that with the increase of trial length complete compensation of hematological and biochemical profiles of rohu might be achieved. The results indicated that the implementation of alternative 7 days low and high protein diet feeding during aquaculture of carps could make economize the operation through minimizing the feed input cost.

A study on the average yield of different species pf prawns used for freezing and canning

The paper deals with the average yield of four spp of prawns viz. Metapenaeus dobsoni, Metapenaeus affinis, Parapenaeopsis stylifera and Penaeus indicus on conversion to peeled and deveined (PD), cooked and peeled (CP) and head less shell on (HL) forms in the different months of a year and the likely variations observed in the average yield.

A statistical study on weight loss in frozen prawns

This paper deals with an extensive study conducted to estimate the extent of weight loss in frozen prawns. The weight Joss varied from 7 to 12% in peeled and deveined (PD), 5 to 7% in headless (HL) and about 7% in cooked and peeled (CP) prawns from the date of processing to the date of inspection, normally within two weeks. To compensate the weight loss nearly 11% of excess material is being added with every frozen block resulting in an average annual loss of Rs. 2.68 crores in foreign exchange. The relevant data pertain to the period 1971 to 1973 and the annual average loss was estimated for the ten years ending 1973.

Genetic diversity of cattle in South China as revealed by blood protein electrophoresis

Genetic variation of 31 blood protein loci in 236 cattle from eight South China populations (including mithan, Bos frontalis) and a Holstein population was investigated by means of horizontal starch gel electrophoresis. Thirteen loci (ALB, CAR, Hb-b, Np, PGM, Amy-I, PEP-B, AKP, 6PGD, Cp, Pa, EsD, and TF) were found to be polymorphic. The comparison of average heterozygosities (H) shows that all the native cattle embrace a rich genetic diversity Our results on protein polymorphism suggest that cattle in China originated mainly from Bos indicus and Bos taurus; Xuwen, Hainan, Wenshan, and Dehong cattle and the Dehong zebu are close to zebu-type cattle, and Diqing and Zhaotong cattle are close to the taurine. The mithan was very different from other native cattle, and we suggest that its origin was complicated and may be influenced by other cattle species.

Effect of diets containing various protein and energy levels on some growth parameters, survival and the production of Astacus leptodactylus in different salinities

A factorial experiment was conducted for 60 days to determine of the response of Narrow clawed crayfish Astacus leptodactylus (average weight of 17±2.3 g) to diets containing various protein and energy levels. Nine diets containing three levels of protein (30, 35 and 40 %) and three levels of energy (300,370 and 450 kcal/100g) were formulated and prepared in this trial. Each diet also was used in two levels of salinity include 0 (fresh water) and 12 ppt(Caspian sea water). So this study was conducted with 18 treatments and triplicates random group of 5 crayfish per each 110-litre tank. Weight Gain, Feed conversion ratio (FCR), Protein Efficiency Ratio (PER), Net Protein Utilization (NPU), Daily Food Consumption (DFC), Survival (SVR) and body composition of tail-muscle meat of animal were determined. Comparing the growth parameters in response to interaction between protein, energy and salinity levels demonstrated that all growth parameters have difference between them significantly (p<0.05). Comparing between survival in fresh and Caspian Sea water showed difference significantly. Compare the body composition results indicate the greatest amount of protein absorption in diet number 2(30/370) on fresh water condition. Results from this study indicate that narrow clawed crayfish can be fed a practical diet containing 30% protein and 370 Kcal/100g on non-salinity water which is the optimize CP percentage for their producer’s profits.