种子超干保存种质的研究

当今，生物资源——“物种”正在以地质史上前所未有的速度灭绝或走向濒危。保护生物多样性已是世界范围内环境保护和生物学研究的热点。基于种子具有完善的保护结构和内潜的隐蔽生命以适应逆境的微妙机制，种子库被认为是保护植物种质资源的主要措施。现阶段世界各国的种子库均为低温库或超低温库，可观的建库投资和运转费用的经济负担直接影响到贫困国家和地区植物种质资源及时有效的保护，寻求取代低温库的廉价贮藏种子的策略和技术已列为联合国FAO／IBPGR的全球战略措施。 此乃本文的工作背景和最终目标。早期的研究已查明，通过降低种子含水量可以在适当提高温度的条件下达到在较高含水量在低温下同样的贮藏效果，但是由于干燥技术以及5%为种子安全含水量下限这一经典概念的局限，种子贮藏的含水量一直没有突破5%下限。超干贮藏即以此为依据，摸索以适当的干燥技术将种子含水量降至这一经典下限以下，即采取超低含水量种子密封贮藏的方法，以达到部分或完全取代低温库的目的。 作为种子贮藏的方法首先要保证种子活力的保持。本文选用了不同化学组份以及不同粒度大小的种子为材料，涉及到40个不同的种和品种，针对不同类型的种子摸索了适宜的干燥方法及辅助措施，从种子生活力和活力、细胞形态结构、细胞器发育和功能以及若干生理活性指标等多元角度上确认种子含水量可以安全降至5%以下，除了个别敏感类型种子外，在常温条件下能大幅度增强种子的耐藏性。 本文对种子耐干性差异以及辅助干燥措施提高种子耐干性进行了水分热力学方面的分析，认为种子本身的特性决定了种子内部水分吸附位点对关键水分束缚力量的不同，导致在相同干燥过程中失去关键性水分程度的不同，从而造成种子在耐干程度上有所差异。干燥的辅助措施可以通过改变种子中的化学成份增强种子对关键性水分吸附的力量，继而使得关键性水分不会轻易丢失。超干延缓种子劣变的原因在于，MC不能真实反映种子水分水平，一定MC范围内的超干种子的关键性水分仍然存在，即使在极度干燥的种子中，虽然一部分束缚水已失去，但对导致和抑制脂质过氧化的正负效应折衷的结果使劣变反应从整体上还是受到抑制。内源抗氧剂的活性及含量以及劣变产物MDA的积累量表明种子在超干贮藏过程中脂质过氧化被部分抑制，酶类抗氧剂系统保存完好，非酶类抗氧剂被消耗的量减少，使得种子在结束贮藏进入萌发状态时仍具有较高水平的自由基清除系统的正常运转能力。无氧贮藏，和外源抗氧防护等预处理措施使种子在超干贮藏过程中能更好地清除活性氧的伤害，吸湿回干、PEG等处理使超于种子吸胀时膜系统功能及结构更具完整性。回水预处理诱导的生化修补效应可保留于种子内部，以便在种子再次吸水时及早发挥作用。本文已在干前及干后预处理辅助措施上找到了若干行之有效的方法，其中有些是首次应用于种子技术范畴。 本文在总结系列试验研究结果和纵观近年来国内外在此领域的动态后，认为一般常规型种子在适度超低水分状态下不仅自身不受脂质过氧化的危害，并能保持种子内外形态学和种质遗传性的完整性，而且可使种子内部代谢停顿，更有利于耐藏性的大幅度提高，加上超干技术上的不断完善，为今后建立节能种子库展示了希望。

Microbiological quality of some iced fishes of Imphal market, Manipur

Imphal is the main marketing centre of fish in Manipur. As fish production of the state is not sufficient to meet the demands, about 120 metric tons of iced fishes are annually brought from other states and sold in this market. Microbiological quality of iced Wallago attu, Labeo rohita, L. gonius and Aorichthy aor in respect of total fungal count (TFC), total plate count of bacteria (TPC), Most Probable Number (MPN) of coliforms, Streptococci, Staphylococcus, Salmonella and Escherichia coli in four tissues (skin, muscle, gill and intestine) were analysed. In all cases, the counts were highest in the gills and lowest in the muscles. The values of TFC, TPC, coliforms, Streptococci and Staphylococci were 0-10³/g 10(sup)6-10⁸/g, 2-α/g, 10-10⁵/g, 10-10⁵/g respectively. E. coli and Salmonella were not detected in any of the samples while the ice used in the preservation contained 10⁵-10⁷ of TPC per gram. The microbiological qualities of the iced fishes of Imphal market were adjudged poor. The extremely high counts of bacteria might be due to (1) poor quality and left over fishes being packed, (2) contact with contaminated ice and (3) repeated thawing and freezing during the process of marketing and transportation.

The processing of fish and other marine resources

The preservation of freshness in fish (commonly referred to as fish handling) and the conversion of fresh fish into other products can both be considered as methods of processing. In commercial practice, however, the term "processing" is confined to the conversion of fresh fish into other products.

苦苣苔科植物的组织培养和离体保存

苦苣苔科(Gesneriaceae)植物种类繁多, 全世界约150属3700余种,我国有58属470余种，大部分具有极高的观赏价值，许多是传统的民间草药。虽然我国苦苣苔科植物资源丰富，然而很多种类分布区域狭窄，种群数量稀少，加之生境受到破坏，许多已经面临灭绝的危险。本研究拟通过组织培养、玻璃化超低温保存以及快速繁殖，达到保护和扩繁珍稀濒危苦苣苔科植物的目的。 以药用唇柱苣苔（Chirita medica D. Fang ex W. T. Wang）和粉绿异裂苣苔（Pseudochirita guangxiensis W.T.Wang var. glauca Y. G. Wei et Y. Liu）为材料，取幼嫩叶片为外植体，通过组织培养实验得到最佳诱导不定芽培养基：MS培养基附加30 g l-1蔗糖，7.5 g l-1琼脂，药用唇柱苣苔附加0.10 mg l-1 BA ，0.10 mg l-1 NAA，粉绿异裂苣苔附加0.05 mg l-1IAA，1.00 mg l-1BA。最高不定芽诱导率分别为：90.3%和85.0%。最佳生根培养基：1/2MS培养基附加30 g l-1蔗糖，5 g l-1活性炭，7 g l-1琼脂，生根率为100%，诱导产生6.11条根，根长为18.8mm（药用唇柱苣苔）；1/2MS培养基附加10-20g l-1蔗糖，1 g l-1活性炭，7 g l-1琼脂，诱导生成6.8-7.4条根，均长17.7-22.0mm（粉绿异裂苣苔）。 在组织培养的基础上进行了苦苣苔科植物的玻璃化超低温冷冻保存研究。以烟叶唇柱苣苔（C. heterotricha Merr.）和濒危植物药用唇柱苣苔叶片外植体为材料，经过自然干燥、装载液处理、玻璃化溶液处理、液氮冷冻保存，成功实现了玻璃化超低温冷冻保存，经过液氮冷冻保存后的材料可以继续分化、生长。适当时间的玻璃化试剂处理对于材料无致死作用，不经液氮冷冻，可以达到100%存活。-20 oC 、-40 oC、液氮保存后，存活率随温度下降而下降，表明冷冻致死的原因在于冰晶形成；提高冷冻后成活率的关键是控制干燥脱水，经过适当的自然干燥，材料存活率分别达到50.0%和27.8%。 以叶片为外植体材料，通过组织培养和快速繁殖可以大规模扩繁苦苣苔科植物。主要步骤为：外植体叶片消毒→不定芽诱导培养→生根诱导培养→继代保存或炼苗移栽，经过3-4个月时间可获得大量栽培植株。已成功保存并培养了40余种苦苣苔科植物，包括濒危苦苣苔及高观赏价值苦苣苔。

Avaliaçāo do estado dum recurso e da pesca

One of the key sectors of fisheries research is assessment and exploration which should have as main goal the support to the fishing industry: analyzing the effects of fishing on stocks, determining the potential stock to be exploited and the formulation of rules for an appropriate exploitation and preservation of resources. In this paper the author intends to analyze some aspects of the investigation in support to fishery management.

Sperm cryopreservation of Indian major carp, Labeo rohita: cryodiluents, sperm: cryodiluent dilution ratio and cryoprotectan t concentration

Cryogenic preservation trials of spermatozoa of Labeo rohita were carried out. Twenty four cryodiluents (extender + cryoprotectant), with the combination of six extenders such as egg-yolk citrate, urea-egg-yolk, 0.9% NaCl, Kurokura-2, Ma and Mb and four cryoprotectants viz. DMSO, glycerol, methanol and ethanol, were used to screen out the suitable cryodiluents. Sperm was preserved in 0.25ml plastic straw in programmable freezer. Two step freezing method was followed. Sperm preserved with egg-yolk citrate and urea-egg-yolk containing 10% DMSO showed best post-thaw motility (80%) followed by 0.9% NaCl (60%) and Kurokura-2(30%) solutions. Sperm with the extenders M" and Mb clotted at the time of equilibration and also after few days of preservation. Egg-yolk citrate mixed with ethanol and methanol also showed good percentage of motility (80%) but egg-yolk citrate with glycerol showed less sperm motility (>60%). To determine suitable dilution ratio of milt and cryodiluent two best extender eggyolk citrate and urea-egg-yolk with four cryoprotectants such as DMSO, glycerol, methanol and ethanol at different ratio viz 1:2,1:4,1:7,1:10,1:15 and 1:20 were used. Highest post-thaw motility (>80%) was observed when milt was preserved with egg-yolk citrate containing 10% DMSO at 1:2, 1:4, 1:7 and 1:10 dilutions. Meanwhile using glycerol as cryoprotectants provided less post thaw motility at lower dilution ratio but with the increase of its dilution showed good sperm motility compared with other cryoprotectants. Finally, evaluation on the effect of cryoprotectant concentration on post-thaw sperm motility was conducted. Egg-yolk citrate and four cryoprotectant i.e. DMSO, glycerol, methanol and ethanol with six different concentrations namely 5%,7%, 10%, 15%, 20% and 30%.were evaluated. Among the cryoprotectants DMSO, methanol and ethanol showed highest post-thaw motility (about 80%) at 7% and 10% concentrations. Although glycerol was not suitable at low concentration but its 20% and 30% concentration levels provided best post-thaw motility. No post-thaw motility was obtained with DMSO at 30% concentration. The overall analysis on cryoprotectant concentration indicated that below 5% and above 20% cryoprotectant concentrations could not be suitable for effective cryopreservation of spermatozoa.

Suitability of sodium lactate as cryoprotectant and its effect on gel forming ability and protein denaturation of croaker fish surimi during frozen storage

The effect of sodium lactate is compared with sucrose + sorbitol + sodium tri-poly phosphate as cryoprotectant on gel forming ability & protein denaturation of croaker surimi during frozen storage at -20±2°C for 90 days was evaluated. The quality of Croaker surimi with 6% (w/v) sodium lactate was examined in terms of biochemical parameters of muscle protein, thaw drip, gel strength and calcium ATPase activity :.omparing with those of surimi added with sucrose/sorbitol & without additive as control. Both the cryoprotectants minimized the negative effects of frozen storage on physico-chemical traits of myofibrillar proteins which was evident from the biochemical and sensory parameters. The residual Ca2+ ATPase activity and gel strength of surimi with sodium lactate were higher than those of control throughout 90 days of storage. Ca2+ A TPase activity and gel strength found a high positive correlation. From the results, it was found that sodium lactate was equally effective in preservation of croaker muscle protein native structure during frozen storage as the sucrose/ sorbitol and also less sweet without any risk of maillard browning.

Marketing of small indigenous species of fish (SIS) in three selected fish markets of Mymensingh

A socio-economic survey was conducted round the year in three fish markets at Mymensingh, Bangladesh. The selected markets were categorized as rural market (Sutiakhali market), a peri-urban market (Kamal Ranjeet market, BAU) and an urban market (Notun Bazar market, Mymensingh town). It was learnt from the survey that the availability of Small Indigenous Fish Species (SIS) declined to a great extent over the last few years and at presently many of such fish species are either threatened or at the edge of extinction. The supply of SIS was highest in KR market (37% of total) and more or less similar in Notun Bazar and Sutiakhali fish market (25 and 27% respectively). The total supply of SIS fluctuated from 25% to 35% throughout the year in these markets. About 48 SIS were found in the sampled markets over the survey period. The highest number of species (45) was found in KR market followed by Notun Bazar (42) and Sutiakhali (37) fish markets. During the survey, three critically endangered species namely, schilbid catfish, garua catfish and rita were found in these markets. Beside these, other 11 and 10 species were listed to be endangered and vulnerable respectively. The biodiversity of 21 SIS found in three markets were no threat at all. Three species (guntea loach, Indian glass barb and flying barb) were 'data deficient' as reported by the IUCN Red Book (IUCN-Bangladesh 2000). From the supply point of view small prawn, spotted snakehead, stinging catfish, pool barb, striped dwarf catfish, Gangetic mystus, walking catfish and tank goby were the prominent fish. The least available species found in this survey were lesser spiny eel, barred spiny eel, Gangetic ailia, freshwater garfish, zig-zag eel, flying barb, Ganges river sprat, freshwater river shad and dwarf gourami. The weight of SIS available in Notun bazar was highest and nearly double than other two markets. There was no significant difference recorded in the supply of SIS in Sutiakhali and KR markets. The average monthly SIS supply was 185, 192 and 467 kg in KR, Sutiakhali and Notun Bazar, respectively; therefore, the cumulative average supply was 844 kg per month in three markets. The price of SIS ranged widely from taka 50-450/kg depending on species, location of market, time of purchase and the condition of fish. In general small prawn, ticto barb, dwarf gourami, Gangetic leaffish, and Annandale loach were sold at a lower price (ranged taka 50-100/kg) and these species could be considered at the bottom of the market-price list. Other SIS like walking catfish, climbing parch, butter catfish, cotio and schilbid catfish valued as highest price (ranged taka 150-450/kg). There was no specific marketing chain for SIS in Mymensingh region. The components of marketing channels and their expansion varied with seasons and locations. The general pattern, however, was as this - after buying fish from fish farmer/fishermen, middlemen (locally known as Foria) used to buy fish to wholesale market and sell to the wholesalers. The retailers used to buy fish from wholesaler through auction to the highest bidders. The retailers then send the fish to particular market where the fish reached the consumers. The livelihood strategy of SIS retailers in three fish markets showed that socio-economic constraints such as low income, poor educational background, low economic status and lack of capital are the main constrains [sic]. Most of the retailers proposed that government should control the fish price throughout the year, so that the producers can get reasonable and stable price. Construction of cold storage and preservation facilities at market sites, improvement of road and communication, improvement of physical market facilities and reduction of market chain is essential. Credit facilities, improvement of their standard of living, health and sanitary condition, housing condition, children education and access to drinking water facilities were identified as additional aspects to improve socio-economic condition of SIS retailers.

Behaviour of marine microflora towards antibiotics 1. Sensitivity towards chlortetracycline (CTC)

The sensitivity of 61 cultures of bacteria isolated from fish towards chlortetracycline (CTC) at 5 ppm and 20 ppm levels has been determined on two solid media: sea water agar (SWA) and a distilled water based medium consisting of peptone, beef extract, glucose and NaCI (PBGA). The cultures employed consisted of (i) gram-negative rods of marine origin (Achromobacter, Pseudomonas, Vibrio and Flavobacterium) and (ii) gram positive organisms (Micrococci and Corynebacterium). Depending on the inhibition zone diameter, the order of CTC sensitivity was found to be Pseudomonaspreservation by CTC is discussed.

On the importance of ship-bottom fouling by marine organisms: a techno-economic survey

A high attention has been paid for constant research on the preservation of materials in the marine environment. This includes all phases of design, development, applied engineering and economics which may influence the construction and operation of ships and underwater installations.

Quality loss in prawns due to double freezing

Icing is the practice for preserving prawns on board fishing boats in India. Majority of these boats need to preserve the catch only for a few hours because of the short duration of the fishing trip. However, with the anticipated introduction of a considerable number of bigger fishing vessels which can remain in the fishing ground for longer periods, more than fortnight, preservation methods, other than icing are required to retain prime quality. Freezing and cold storage of whole prawns on board followed by thawing and processing on land is a possible proposition. The extent of quality loss in prawns during these operations is one of the important points to be considered. Hence, laboratory scale studies were undertaken on double freezing of prawns and the results are dealt within this communication.

Canning of Lactarius

Though not a sizable fishery on national level, landings of lactarius (Lactarius lactarius) is considerable in the states of Kerala and Tamil Nadu. Apart from a note (Anon, 1965) no published work is available on its preservation by canning. Fish processed by this method on storage loses its flavour and the lustrous skin is peeled off. The present note describes a method in which these factors also are taken care of.

Preparation of mussel meat by drying

The paper describes a simple and cheap process for the preservation of mussel meat by drying. The method involves blanching the mussel meat shucked from purified live mussels in 5% boiling brine for 5 min followed by drying to moisture of 10 to 15%. The product stored in glass bottles or polythene bags suitably sealed, has a storage life of about six months after which the organoleptic qualities begin to deteriorate. No preservative is used at any stage of processing and the yield of the product is approximately 20%. The major type of spoilage during storage is brown discoloration. Spoilage due to insect infestation is also common unless packed properly.

Ultrastructural mechanical and material characterization of fossilized bone

Bone plays a key role in the paleontological and archeological records and can provide insight into the biology, ecology and the environment of ancient vertebrates. Examination of bone at the tissue level reveals a definitive relationship between nanomechanical properties and the local organic content, mineral content, and microstructural organization. However, it is unclear as to how these properties change following fossilization, or diagenesis, where the organic phase is rapidly removed and the remaining mineral phase is reinforced by the deposition of apatites, calcites, and other minerals. While the process of diagenesis is poorly understood, its outcome clearly results in the potential for dramatic alteration of the mechanical response of biological tissues. In this study, fossilized specimens of mammalian long bones, collected from Colorado and Wyoming, were studied for mechanical variations. Nanoindentation performed in both longitudinal and transverse directions revealed preservation of bone's natural anisotropy as transverse modulus values were consistently smaller than longitudinal values. Additionally, modulus values of fossilized bone from 35.0 to 89.1 GPa increased linearly with logarithm of the sample's age. Future studies will aim to clarify what mechanical and material elements of bone are retained during diagenesis as bone becomes part of the geologic milieu. © 2007 Materials Research Society.

Single UV excitation of Hoechst 33342 and propidium iodide for viability assessment of rhesus monkey spermatozoa using flow cytometry

Many fluorescent probes excited by visible light have been used to assess sperm quality by flow cytometry. Developing a viability evaluation method using UV excited stains would be useful for multiparameter analysis of sperm function. This investigation was conducted to determine the efficacy of Hoechst 33342 (H342) and propidium iodide (PI) dual staining for evaluating rhesus monkey sperm viability through use of flow cytometry and excited by a single UV laser. The results showed that the live cells stained only with H342 strongly correlated with expected sperm viability, and flow cytometric analyses were highly correlated with fluorescence microscopic observation. Using H342/PI/SYBR-14 triple staining method, it was found that the live/dead sperm distributions were completely concordant in both H342/PI and SYBR-14/PI assays. In addition, this dual staining was extended with fluorescein isothiocyanate-conjugated peanut agglutinin (FITC-PNA) to simultaneously analyze viability and acrosome integrity of sperm cryopreserved using two different extenders, TTE and TEST, and indicated that TTE offered better Preservation of plasma and acrosome integrity than TEST Therefore, the H342/PI dual staining provides an accurate technique for evaluating viability of rhesus monkey sperm and should be valuable for multiparameter flow cytometric analysis of sperm function.