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1.
采用细胞松驰素Ｂ（Ｃ．Ｂ）处理法，研究了诱导中华绒螯蟹（Ｅｒｉｏｃｈｅｉｒｓｉｎｅｎｓｉｓ）三倍体、四倍体的可能性和处理条件。实验所用Ｃ．Ｂ浓度为０．５～２．０ｍｇ／Ｌ，处理的起始时间分别在受精后１０～２５ｍｉｎ（诱导三倍体）和８．５～９．８ｈ（诱导四倍体），处理的持续时间为１０～２５ｍｉｎ。三倍体的适宜诱导条件为卵子在受精后１５ｍｉｎ，浓度为１．５ｍｇ／Ｌ的Ｃ．Ｂ溶液处理１８ｍｉｎ。三倍体诱导率最高可达５８．１８％。根据诱导三倍体的适宜Ｃ．Ｂ处理浓度和处理时间，用１．５ｍｇ／ＬＣ．Ｂ溶液处理卵子１８ｍｉｎ，获得了２９．４６％～５７．８９％的四倍体胚胎。另外，还对中华绒螯蟹三倍体、四倍体胚胎的诱导技术、诱导参数对胚胎发育的影响等有关问题进行了分析讨论 相似文献
2.
以江苏文蛤为实验材料,采用50mg／L秋水仙索和400μM6-二甲基氨基嘌呤（6-DMAP）处理、抑制受精卵第一极体排放和抑制第一次卵裂的方法,进行四倍体诱导研究。根据不同处理时间,D形幼虫的孵化率分别为38．6％和34．58％。经胚胎和幼虫发育观察发现,在诱导组中异常原肠胚的比例高于对照组;受精后48h,诱导组的滞育担轮幼虫的比例显著高于对照组。在受精后48h取担轮幼虫制作染色体片,诱导组中四倍体幼虫的比例为7．56％-36．2％和9．2％-28-3％,最高诱导率达至Ⅱ了36．2％。同时每个处理组都发现了不同的非整倍体存在。本研究表明,该项技术能有效地诱发文蛤四倍体幼虫,但面临四倍体幼虫存活率低的问题,今后应设法提高四倍体胚胎和幼虫的发育水平。 相似文献
3.
6—DMAP诱导太平洋牡蛎三倍体——抑制受精卵第二极体释放↑（*） 总被引：24，自引：3，他引：21
于１９９６～１９９７年,用６－ＤＭＡＰ抑制太平洋牡蛎受精卵第二极体的释放,诱导产生三倍体,最高诱导率为９３．８％。该实验组胚胎孵化率为８１．０％,Ｄ形幼虫畸形率为１７．６％。实验选用Ｌ９（３４）设计,进行三因素三水平的正交试验：６－ＤＭＡＰ浓度,设３００、４５０和６００μｍｏｌ／Ｌ等三水平;诱导时机（观察静止状态的受精卵,以受精卵出现第一极体的百分率指示）,设１０％、３０％和５０％等三水平;诱导持续时间,设１０、１５和２０ｍｉｎ等三水平,试验设三次重复。解剖法获取精卵,人工授精。染色体倍性检查采用染色体计数法。根据正交试验直观分析结果,得出诱导太平洋牡蛎三倍体各因素的最优水平组合：当３０％受精卵出现第一极体时,将受精卵浸泡在含６－ＤＭＡＰ４５０μｍｏｌ／Ｌ的海水中１０ｍｉｎ;三因素的主次顺序：６－ＤＭＡＰ浓度→诱导时机→诱导持续时间。６－ＤＭＡＰ的浓度和诱导时机对三倍体诱导率的影响显著;但诱导持续时间影响不显著。部分试验组得到４．２％～１３．６％的四倍体和１０％左右的非整倍体,这种情况可能与受精不同步抑制第一极体释放而导致的染色体多极分离有关。 相似文献
4.
用细胞松弛素B（CB）处理九孔鲍♂×盘鲍♀受精卵,分别抑制其第一极体和第二极体、以及第二极体释放诱导异源三倍体。水温24℃,九孔鲍♂×盘鲍♀授精后10min,用浓度0.6～1.0mg/L的CB持续处理受精卵20～25min,抑制其第一极体的排放。而九孔鲍♂×盘鲍♀在受精后27min,当40%～50%受精卵排出第一极体时,用浓度0.6～1.0mg/L的CB持续处理受精卵10～15min,分别统计对照组和药物处理组的担轮幼虫率,并用倍体分析仪检测各组稚鲍的倍性。结果表明：对照组和药物处理组担轮幼虫的倍性较复杂,起始处理时间为10min,CB药物处理浓度为0.6mg/L,持续处理时间为25min,其三倍体率可达40.67%,担轮幼虫的孵化率为26.44%。起始处理时间为27min,CB药物处理浓度0.6mg/L,处理持续时间为10min,其三倍体率可达48.11%,担轮幼虫率的孵化率为29.86%。 相似文献
5.
以热休克抑制受精卵第一次卵裂，进行日本沼虾四倍体诱导实验，热休克温度在３８、３９、４０和４１℃，于产卵２１０￣２３０分钟后，处理１￣２分钟均可获得四倍体胚胎，４０℃处理１．５分钟，四倍体胚胎诱导率达３６．８％，３８和３９℃处理２分钟后所获得的嵌合体胚胎比率大于四倍胚胎比率，而热休克温度４０℃时，四倍体率显著高于合体比率，４１℃处理时由于有丝分裂异常导致胚胎死亡率高，更适宜的处理时间有待进一步研究， 相似文献
6.
7.
6_二甲基氨基嘌呤诱导栉孔扇贝三倍体 总被引：14，自引：1，他引：13
用６－二甲基氨基嘌呤（６－ＤＭＡＰ）处理以抑制栉孔扇贝第二极体释放产生三倍体的适宜诱导参数进行了试验研究,结合ＤＡＰＩ染色和荧光显微镜观察,分析了开始处理前和处理结束时的受精卵染色体核相组成与三倍体诱导率和幼虫成活率的关系,对三倍体处理组和对照组幼虫的生长进行了观察,结果表明,获得充分成熟的卵子,起始处理时间以第一极体占４０％,药物浓度６０ｍｇ／Ｌ,持续处理时间１５ｍｉｎ及产卵至处理结束保持水温２ 相似文献
8.
用1500μW/cm2的紫外线照射长牡蛎(Crassostrea gigas)精子60 s以进行灭活处理,并使之与栉孔扇贝(Chlamys farreri)卵子受精,在卵子受精后排出第一极体前用6-DMAP(50 mg/L)处理受精卵,持续处理35 min,抑制第一极体和第二极体的排放,诱导异精雌核发育四倍体。采用二脒基苯基吲哚(DAPI)荧光染色显微观察法,对灭活的长牡蛎精子诱导的栉孔扇贝雌核发育四倍体早期胚胎发育过程进行细胞学观察。结果表明:经紫外线灭活过的长牡蛎精子进入栉孔扇贝卵子后发生轻微膨胀;在第一次卵裂中期,精核形成一致密的染色质小体(DCB),位于两组分开的母本染色体之间,不参与核分裂;第一次卵裂结束时DCB滞留于两卵裂球的分裂沟上或进入其中一分裂球中;第二次卵裂过程中,DCB的去向与第一次卵裂时基本一致。6-DMAP处理有效地抑制了第一极体和第二极体的排出,从而使雌核四倍化。对担轮幼虫染色体倍性分析结果表明,通过本方法可以获得6.25%的四倍体幼虫。本研究还对灭活的异源长牡蛎精子诱导栉孔扇贝雌核发育四倍体过程中产生的复杂倍性、核物质分离紊乱及多精附卵现象进行了观察和分析。 相似文献
9.
冷、热休克法诱导黄颡鱼三倍体的比较研究 总被引：5，自引：0，他引：5
分别采用冷、热休克抑制第二极体释放的方法诱导黄颡鱼三倍体。结果表明,在卵受精后2min,5℃处理20min,胚胎时期的三倍体率达70%左右,孵化率50%左右,幼鱼时期三倍体(含嵌合体)的检出率为25%,此条件为冷休克处理的优化参数;在卵受精后2min,40℃处理2min,胚胎时期的三倍体诱导率达58%,孵化率为39%,幼鱼时期三倍体(含嵌合体)的检出率为40%,此条件为热休克处理的优化参数。正交分析得出,冷休克条件下起始休克时间是原肠期三倍化率和孵化率的重要影响因子,温度对畸形个体的产生有重要影响;热休克条件下,参考三倍体率、畸形率、孵化期相对存活率三者而言,休克温度均是重要因素。比较观察到冷休克处理组的胚胎受损情况严重,后期的成活率较热休克处理组要低,总体诱导效果逊于热休克处理组。 相似文献
10.
为研究杂交鲍的三倍体诱导技术,通过预试验,观察和了解23.5 ℃下杂交鲍受精卵第一极体和第二极体释放的时间。采用细胞松弛素B抑制第二极体释放的方法,分别在不同的诱导起始时间、诱导剂浓度及诱导持续时间下抑制第二极体的释放,测定鲍的三倍体率,进行了杂交鲍三倍体的诱导试验。试验结果表明:杂交鲍在水温23.5 ℃下,处理起始时间为20 min,诱导剂浓度为0.70 mg·L-1,诱导的持续时间为10 min时,表观三倍体率最高,达75 %以上。检查稚鲍的倍性,三倍体率达到66 %。 相似文献
11.
The triploid technology is a new frontier in shellfish aquaculture and has shown encouraging results in numerous shellfish species. We induced triploid larvae in the Yesso scallop, Patinopecten yessoensis, using hyperosmotic shock for the first time in this study. Different induction parameters, including salinity strength, treatment starting point, and the duration, were tested. The highest triploid ratio of D‐shaped larvae (72.12%) was obtained by 60 ppt salinity treatment for 20 min at the first appearance of zygote showing polar body II (PB2). A significantly faster growth rate was observed during the swimming larvae stage despite a decrease in the hatching and survival ratios. The triploid ratio decreased to 46.67%, and approximately 1.76 million triploid juveniles were harvested after 90 days of cultivation. The treatment parameters can be further optimized to improve the yield of Yesso scallop triploids. 相似文献
12.
Triploid induction in Australian greenlip abalone, Haliotis laevigata (Donovan), was conducted by blocking the formation of the second polar body using cytochalasin B (CB). Twenty minutes after fertilization, the zygotes of greenlip abalone were treated with four CB concentrations (0, 0.25, 0.5 and 0.75 mg L−1) for 10, 15 and 20 min. The ploidy of resultant larvae was determined using flow cytometry at 72-h post fertilization. Our study showed that fertilization, hatching, survival and induced triploidy of abalone larvae were significantly affected by the CB concentration and treatment duration. The effective range of CB concentration for triploid induction on greenlip abalone was 0.5–0.75 mg L−1 with an induction duration of 10–15 min. The results indicate that the most effective treatment combination for triploid induction in greenlip abalone is 0.5 mg CB L−1 for 15 min starting at 20-min post fertilization. 相似文献
13.
Different durations of a cytochalasin B (CB) treatment were tested for tetraploid induction by meiosis inhibition in the dwarf surfclam Mulinia lateralis Say. Cytochalasin B, 0.67 mg L?1, was applied to newly fertilized eggs at 8–10‐min post fertilization and removed when in the untreated eggs: (1) Polar body 1 (PB1) was released in 90% of the eggs and polar body 2 (PB2) began to form (T1); (2) Polar body 2 was released in about 25–30% of the eggs (T2); (3) Polar body 2 was released in about 70–75% of the eggs (T3); or (4) eggs began to enter mitosis I or the polar lobe began to form (T4). Three replicates were produced using different sets of parents. The ploidy of resultant larvae and juveniles was determined by flow cytometry. Blocking PB1 alone in T1 groups produced mostly tetraploids, and longer CB treatments in T2 and T3 resulted in increasing numbers of pentaploids. In T4 groups where both PB1 and PB2 were inhibited, larvae were predominantly pentaploids. Pentaploid larvae were arrested at the trochophore stage. The majority of tetraploid larvae died as trochophores, although a small fraction reached D‐stage. Among 478 juvenile clams sampled from a T1 group, three (0.6%) were confirmed as tetraploids. This study shows that tetraploid embryos can be produced at high efficiencies (40–90%) by blocking meiosis I. Tetraploids produced by meiosis inhibition in normal eggs are viable in M. lateralis, but their survival beyond metamorphosis is extremely low. 相似文献
14.
为探讨大黄鱼(Larimichthys crocea)和黄姑鱼(Nibea albiflora)精子的紫外辐射灭活适宜剂量及其激活大黄鱼卵子发育为胚胎的效果,以Ringer氏液为稀释液,按1:30稀释大黄鱼和黄姑鱼精子,采用自制紫外灭活装置[紫外辐射强度2200μW/(cm~2·s),紫外波长254 nm]对这2种鱼的精子进行紫外照射处理及活力测定,然后与正常大黄鱼卵子进行人工授精,授精后一部分卵未作冷休克处理,另一部分卵进行了冷休克处理(受精2 min 30 s,3℃海水,冷休克10 min),并进行了早期胚胎成活率和仔鱼孵化率的测定与比较。结果显示:1)大黄鱼和黄姑鱼精子的激活率与紫外照射处理时间呈负相关,精子的快速运动时间变化呈典型的Hertwig效应。2)未冷休克组中大黄鱼和黄姑鱼诱导的早期胚胎成活率与精子的紫外照射时间总体呈负相关,而仔鱼孵化率呈Hertwig效应。3)冷休克组中大黄鱼和黄姑鱼精子诱导的早期胚胎成活率和仔鱼孵化率随紫外照射时间的增加呈Hertwig效应,分别于2 min 20 s和1min 30 s时达相对峰值,此时,大黄鱼早期胚胎成活率和仔鱼孵化率分别为(38.3±4.3)%和(66.5±5.1)%,黄姑鱼早期胚胎成活率和仔鱼孵化率分别为(43.3±3.3)%和(67.7±6.3)%。分析认为,大黄鱼和黄姑鱼精子遗传失活的紫外辐射剂量分别以308 m J/cm~2和198 m J/cm~2为佳。本研究旨在为大黄鱼雌核发育技术的改进提供依据。 相似文献
15.
三倍体九孔鲍的育种和养成技术研究 总被引：8，自引：1，他引：8
当 5 0 %九孔鲍受精卵出现第一极体后 ,以 6 -DMAP为诱导剂 ,用不同的浓度和不同的持续时间抑制第二极体的释放诱导三倍体 ;并进行了生产性苗种培育和养成试验。结果表明 ,6 -DMAP的诱导浓度为30 0μmol·dm-3 、诱导持续时间为 10min时 ,其胚胎的三倍体率为 90 %以上 ,并且胚胎的孵化率高达 85 % ,幼体的畸形率较低 ,为 5 0 %～ 5 5 % ;经过 7个月培育 ,试验组养成鲍的三倍体率为 6 5 %以上 ;试验组养成鲍的平均壳长比对照组增长 10 % ,平均体重比对照组增重 30 %。与二倍体鲍相比较 ,三倍体鲍在壳长增长和体重增重等方面 ,显示了显著的优势 [t>t0 .0 1( 58) ]。 相似文献
16.
Yi-ping Shen Xi-yuan Zhang Hai-ping He Li-jun Ma 《Aquaculture (Amsterdam, Netherlands)》1993,110(3-4):221-227
Triploidy of the pearl oyster, Pinctada martensii Dunker, was induced with hydrostatic pressure shock. The efficiency of inductions changed with the time of initiation of treatment, the duration of treatment and the intensity of pressure. The most effective procedure for inducing triploidy was the treatment of eggs at 200–250 kg/cm2 5–7 min (for the first polar body retention) and 17–19 min (for the second polar body retention) after insemination for 10 min duration, which resulted in 76% triploid embryos. Under this optimal treatment, high hatchability was also obtained. 相似文献
17.
Large yellow croaker, Pseudosciaena crocea, exhibit sexually dimorphic growth, with females growing faster and reaching larger adult sizes than males. Thus, development
of techniques for preferentially producing females is necessary to optimize production of these species. We have established
a protocol to produce all-female croaker P. crocea through induction of meiotic gynogenesis with homologous sperm. The first set of experiments investigated the ultra-violet
(UV) irradiation on sperm motility and duration of sperm activity to determine the optimal UV dosage for genetic inactivation
of sperm, yet retaining adequate motility for activation of eggs. Milt from several males was diluted 1:100 with Ringer’s
solution and UV irradiated with doses ranging from 0–150 J cm−2. The results indicated that motility and duration of activity generally decreased with increased UV doses. At UV doses greater
than 105 J cm−2, after fertilization, motility was <10% and fertilization rates were significantly lower. Highest hatching rate was obtained
at 75 J cm−2. A second set of experiments was carried out to determine appropriate conditions of cold shock for retention of the 2nd polar
body in P. crocea eggs after fertilization with UV-inactivated sperm by altering the timing, temperature and duration of shock. At 20°C, shock
applied at 3 min after fertilization resulted in higher survival rate of larvae at 6 h after hatching. Results of different
combinations of three shock temperatures (2°C, 3°C or 4°C) and five shock durations (4 min, 8 min, 12 min, 16 min or 20 min)
at 3 min after fertilization demonstrated that shocks of 12 min gave highest production of diploid gynogens. Statistical analysis
revealed that maximum production of diploid gynogens (44.55 ± 2.99%) were obtained at 3°C. The results of this study indicate
that the use of UV-irradiated homologous sperm for activation of P. crocea eggs and cold shock for polar body retention is an effective method for producing gynogenetic offspring. 相似文献
18.
饥饿对白甲鱼(Onychostoma sima)仔鱼摄食、生长的影响 总被引：2，自引：0，他引：2
在实验室条件下,采用观察、测量记录外部形态特征等方法研究了饥饿胁迫下白甲鱼(Onychostoma sima)仔鱼的摄食、生长、形态和存活的变化。结果显示:在水温(20±1.5)℃时,饥饿组和投喂组白甲鱼仔鱼在孵化后第4天开始摄食,初次摄食率为48%,第6天初次摄食比率达到100%。饥饿组仔鱼抵达饥饿不可逆点(PNR)的时间为13.5 d,50%累计死亡率出现时间基本上与PNR出现时间相同(13.5 d),表明第13.5天为白甲鱼早期发育中较为敏感的阶段。试验期间,饥饿仔鱼体重、体长、眼间距和体高分别在第6、7、11和12天后均显著低于对照组(P<0.05)。试验结束(16 d)时饥饿组仔鱼体重、体长和眼径的瞬时增长率均显著低于摄食组(P<0.05),而头长、体高和眼间距差异不显著(P>0.05)。结果表明饥饿对仔鱼生长发育起延迟作用,混合营养期延迟投喂,仔鱼生长发育水平明显低于正常投喂仔鱼。 相似文献
19.
Awatef Trabelsi Jean‐Noël Gardeur Fabrice Teletchea Jean Brun‐Bellut Pascal Fontaine 《Aquaculture Research》2013,44(4):657-666
Based on the analysis of 11 morphometric variables of body (total length, body area and perimeter, myotome height and eye diameter) and yolk sac (length, height, area, perimeter and volume) of pike larvae, the aim of this study was to evaluate how larval size at hatching and growth of larvae hatched from single egg batches vary according to three hatching times: early, mid and late. Hatching time structures strongly pike larval morphometrics. Early hatched larvae have smaller body sizes at hatch, faster growth and higher yolk use efficiency than late hatched ones. Early hatched larvae seem to be premature and hatch at precocious developmental stage whereas late hatched individuals continue their growth within the egg shell and hatch at larger size but with lower reserves (yolk). A compensatory growth phase was observed for the early hatching pike larvae particularly during the first 5 days post hatch. Consequently, no significant difference in body parameters was recorded from day 10 post hatching whatever the hatching time. The higher growth accomplished by early hatched larvae may be related to a particular metabolic activity that converts more efficiently yolk into body tissues. 相似文献