银条茎尖培养快繁及离体根状茎的诱导

 以银条‘二细一粗’品种为试材, 研究了蔗糖浓度、激素组合对茎尖培养和快速繁殖的影响及温度、光照、蔗糖浓度等因素对根状茎离体诱导的影响。结果表明: 茎尖培养较理想的培养基为MS + 蔗糖4 % + 6-BA 0.5 mg·L ^{- 1} + NAA 0.1 mg·L ^{- 1} , 成苗率可达74.0 %。茎尖增殖较适培养基为MS + 6-BA 3.0 mg·L ^{- 1} + NAA 0.5 mg·L ^{- 1} , 每个外植体平均可产生5.6 个芽。根状茎诱导以MS + 蔗糖10 % + 6-BA 510 mg·L ^{- 1}+ CCC 500 mg·L ^{- 1}培养基, 20 ℃全黑暗培养效果最好。     

银条茎尖玻璃化法超低温保存及其植株再生

 为长期稳定保存银条种质, 建立了玻璃化法超低温保存其茎尖的技术体系。选择继代4次的 银条无菌壮苗, 5 ℃低温驯化14 d; 剥取5 mm的茎尖, 在含有0.5 mol·L ^{- 1}蔗糖的MS (无Ca^{2 +} ) 液体培养基预培养1 d; 25 ℃条件下经改良MS + 2PVS₂装载20 min; 在- 20 ℃, 95%乙醇浴中以PVS₂脱水处理4h; 更换新鲜的PVS₃后投入液氮, 保存24 h后取出冷冻管在40 ℃水浴中化冻1 min; 以含有1.2 mol·L ^{- 1}蔗糖的改良MS (无Ca^{2 +} ) 溶液洗涤2 次, 每次10 min; 冻存后的茎尖相对存活率超过70%。将冻存后的茎尖转接到再生培养基上, 暗培养20 d后转入正常光照条件下培养, 存活率达63.7%; 继续培养得到正常分化和生长的再生植株, 生根后可移栽成活。     

Reproductive biology of the commercial sea cucumber <Emphasis Type="Italic">Holothuria spinifera</Emphasis> (Echinodermata: Holothuroidea) from Tuticorin,Tamil Nadu,India

The annual reproductive cycle of the commercial sea cucumber Holothuria spinifera was studied in Tuticorin, Tamil Nadu, India, from September 2000 to October 2001, by macroscopic and microscopic examination of gonad tubule, gonad index and histology of gametogenic stages, to determine the spawning pattern. The gonad consists of long tubules with uniform development. It does not confirm the progressive tubule recruitment model described for other holothurians. The maximum percentage of mature animals, gonad and fecundity indices, tubule length and diameter, with the observations on gonad histology, ascertained that H. spinifera had the peak gametogenic activity during September and October 2001 followed by a prolonged spawning period from November 2000–March 2001.     

Effects of temperature, salinity and pH on larval growth, survival and development of the sea cucumber Holothuria spinifera Theel

For large-scale seed production of sea cucumbers through a hatchery system, it is imperative to know the effects of environmental parameters on larval rearing. Auricularia larvae (48 h post-fertilization) were obtained from induced spawning of Holothuria spinifera and used in experiments to ascertain the effects of temperature, salinity and pH on the growth and survivorship of the larvae. The larvae were reared for 12 days at temperatures of 20, 25, 28 and 32 °C; salinities of 15, 20, 25, 30, 35 and 40 ppt; and pH of 6.5, 7.0, 7.5, 7.8, 8.0, 8.5 and 9.0. The highest survivorship and growth rate and fastest development of auricularia indicated that water temperature of 28–32 °C, salinity of 35 ppt and pH of 7.8 were the most suitable conditions for rearing larvae of H. spinifera.     

Cytotoxic and Apoptosis-Inducing Activity of Triterpene Glycosides from Holothuria scabra and Cucumaria frondosa against HepG2 Cells

The cytotoxic effects of thirteen triterpene glycosides from Holothuria scabra Jaeger and Cucumaria frondosa Gunnerus (Holothuroidea) against four human cell lines were detected and their cytotoxicity-structure relationships were established. The apoptosis-inducing activity of a more potent glycoside echinoside A (1) in HepG2 cells was further investigated by determining its effect on the morphology, mitochondrial transmembrane potential (Δψ_m) and mRNA expression levels of the apoptosis-related genes. The results showed that the number of glycosyl residues in sugar chains and the side chain of aglycone could affect their cytotoxicity towards tumor cells and selective cytotoxicity. 1 significantly inhibited cell viability and induced apoptosis in HepG2 cells. 1 also markedly decreased the Δψ_m and Bcl-2/Bax mRNA express ratio, and up-regulated the mRNA expression levels of Caspase-3, Caspase-8 and Caspase-9 in HepG2 cells. Therefore, 1 induced apoptosis in HepG2 cells through both intrinsic and extrinsic pathway. These findings could potentially promote the usage of these glycosides as leading compounds for developing new antitumor drugs.     

白沙参苗种中间培育关键参数研究

为了解决白沙参参苗个体小、抗逆性及抵御敌害生物的能力弱、成活率低、不利于增养殖的缺陷,研究了白沙参室内水泥池中间培育过程中的关键参数,并在最优条件下进行室内水泥池和室外池塘网箱的培苗,试验结束时测量参苗的体长和体重,计算成活率.结果表明,室内水泥池培育的关键技术参数如下:水温25～30℃,盐度25‰～35‰,培育密度2000～3000头/m2,光照强度2 000 lx.在上述最佳条件下,将体长0.3～0.5 cm、体重0.004 g的参苗16万尾,置于室内水泥池培育1个月后,参苗规格平均达到0.8～1 cm,存活率达到50％以上.然后将其转到室外网箱培养,更换网箱规格并及时疏苗,培育40 d时,参苗规格平均达到3 cm左右,存活率达到60％.     

不同激素配比对离体地灵(Stachys floridana Schuttl.ex Benth)芽增殖的影响

研究了不同激素单独或混合使用对地灵芽增殖的影响。结果表明,6-BA与KT配合使用可诱导茎段产生大量不定芽,但不定芽聚集短缩,难以进一步发育成芽苗。单独使用低质量浓度(0.5mg/L以下)6-BA或KT能够使地灵茎段得到有效增殖,其中KT的效果优于6-BA,KT对试管芽增殖影响比6-BA温和,KT1.5mg/L时仍未出现玻璃化现象。生长素与细胞分裂素混合使用时能够明显促进芽苗的生长,在含有0.5mg/LIAA的MB培养基中添加1.0～1.5mg/L的KT,增殖系数达4.20～4.67,为地灵芽增殖的有效激素质量浓度配比。     

不同支持物对地灵(Stachys floridana Schuttl.ex Benth)试管苗生根的影响

以脱脂棉、琼脂、蛭石、河沙、壤土为支持物,在1/2MS+NAA1.0mg/L+6-BA0.5mg/L培养基上对地灵试管苗生根情况进行了分析。结果表明,支持物对地灵的生根有明显影响,在珍珠岩、琼脂和脱脂棉中地灵的生根率均达到100%,各处理之间差异不显著;生根质量以珍珠岩中最优,幼根能产生大量多级分枝,形成较为强大的根系,且不同支持物试管苗的移栽结果也表明,珍珠岩在地灵试管苗生根中是琼脂的理想替代物。     

Preliminary assessment of large‐scale co‐culture of sandfish (Holothuria scabra) with the Babylon snail (Babylonia areolata) in earthen ponds and in raceways

Holothuria scabra (sandfish) and Babylonia areolata were trialed in two large‐scale co‐culture experiments. Experiment 1 assessed co‐culture in 4 × 400 m² earthen ponds where Babylonia were cultured in a central pen at a density of 400 individuals/m² with sandfish occupying the remaining pond space at 1.1 individuals/m². Sandfish grew from 18.20 ± 6.67 g to 119.03 ± 17.74 g in 92 days and Babylonia (fed trash fish in both experiments) grew from 0.90 ± 0.38 g to 4.93 ± 1.44 g, but Babylonia growth was not increased in co‐culture compared to monoculture. Water and sediment quality varied between co‐culture and monoculture ponds. Neither showed clear improvement due to sandfish culture. Experiment 2 compared non‐segregated sandfish‐Babylonia co‐culture with Babylonia monoculture in 20 m² concrete raceways. Sandfish were cultured at 2 individuals/m² and Babylonia at 300 individuals/m². Sandfish grew up to 1.91 g.day^?1 with 100% survival. Babylonia weight gain was significantly greater in co‐culture raceways (3.35 ± 0.64 g over 61 days), which was double that of Babylonia in monoculture. Substrate total N was reduced by 20% in co‐culture compared with monoculture (p = .032). This provisional study of commercial scale sandfish‐Babylonia co‐culture demonstrates culture compatibility, providing a basis for further system development.     

Food sources,digestive efficiency and resource allocation in the sea cucumber Holothuria forskali (Echinodermata: Holothuroidea): Insights from pigments and fatty acids

Various research projects in Europe and North Africa have recently intended to breed temperate holothurians to alleviate fishing pressure on natural populations. However, to date little is known about the nutritional requirements of East Atlantic and Mediterranean species. In this study, we propose a “natural population”‐oriented approach to characterize food sources, digestive efficiency and resources allocation based on the composition of pigments and fatty acids (FA) in gut contents and tissues (muscles, gonads and digestive tract walls) of wild individuals of the species Holothuria (Panningothuria) forskali (Delle Chiaje, 1823) sampled in Brittany (France). Our study reveals that neither green nor red algae enter the diet of H. forskali in spring and that the only fresh vegetal material found in gut contents is brown algae (very likely diatoms). The high nutritional quality of gut contents however contrasts with the detrital nature of the ingested food sources, suggesting that a trophic upgrading of organic matter occurs before digestion. In addition, unusual FA (i.e. only present in a few groups of living species) such as long‐chain monounsaturated FA (especially the FA 23:1ω9) were found in large proportions in muscles and gonad and their effect on sea cucumber fitness needs further investigation.