黑斑口虾蛄的繁殖生物学研究

【作者】 王春琳；

【导师】 姜乃澄；

【作者基本信息】 浙江大学 ， 遗传学， 2001， 硕士

【摘要】 本文对黑斑口虾蛄(Oratosquilla kempi)雌雄性生殖腺的解剖学、组织细胞学、幼体发生和人工繁殖进行了研究。这一研究对于黑斑口虾蛄生殖腺发育、幼体发生等繁殖生物学和产业化人工育苗方面均有较大的理论和实际意义。现将主要研究结果报告如下： 1、卵巢发育的第Ⅰ期，外观呈透明线状，无色素分布，粘贴于消化道上，卵原细胞附于生殖上皮或其附近的卵巢基质中。第Ⅱ期卵巢始行发育，外观呈灰黄色至浅黄色带状，并向两侧扩展，卵黄形成前期的初级卵母细胞体积开始增大，已有少量卵黄物质出现。第Ⅲ期为初级卵母细胞的卵黄形成期，卵巢外观由浅黄色向橙黄色转变，已可见清晰的卵粒。第Ⅳ期为卵黄形成后期初级卵母细胞，卵巢外观极度膨大，卵子在卵巢内成离散状。研究发现黑斑口虾蛄卵巢发育的特点是卵原细胞可从卵巢边缘生殖上皮深入卵巢中心形成S形增殖区，与其他甲壳动物卵巢有所不同。同时卵原细胞阶段性增殖延续时间较长，至卵黄形成期后期才完全结束。这是导致黑斑口虾蛄产卵延续期较其他甲壳动物更长的重要原因。 2、卵巢发育过程中，性细胞和核的大小，及其核质比在卵巢发育各期区别明显，其中Ⅰ期至Ⅱ期是细胞核增长最明显的时期，但从Ⅲ期早期开始，性细胞体积的增长明显快于核的增长，因此黑斑口虾蛄卵巢发育从第Ⅲ期早期开始进入其营养物质——卵黄形成和贮存的关键时期。故人工繁殖中黑斑口虾蛄亲体培育的重点应抓好卵巢发育早期和中期，即从卵原细胞期到卵黄形成 早期的亲体培育管理。 3、黑斑口虾姑雄性生殖系统由 ＊”字型精巢、类输精管 （deferensoid）、雄性生殖孔、交接器及附属腺组成。精巢外观 呈长“V”字型，与其他甲壳动物呈“H”型精巢明显不同．组织 学观察发现该虾姑输精管壁缺少肌细胞，明显与具肌细胞的输精 管不同，故我们称其为“类输精管”．组织切片发现它的整个精 巢各段均进行着精子的发生，由精巢基膜向管腔依次排列着精原 细胞、初级精母细胞、次级精母细胞和精细胞，且从精巢末段到 类输精管的精巢腔中性细胞渐趋成熟；在类输精管和交接器中仅 为分化中的精细胞，并无精母细胞。各期雄性生殖细胞大小与相 对应的雌性生殖细胞排序相反。根据交接器中的精细胞电镜切片 推测，黑斑口虾姑雄体性成熟后排出的是精细胞，且可随时排放， 而精细胞向精子的变态过程可能在雌体中进行。 一、黑斑口虾姑的生物学最小型雌性为 8．3cm、雄性为 8．ocm． 水温高于15℃的秋末或翌年春季为其交配季节；卵巢正常发育的 水温为 15－3 0 C．卵巢成熟系数以 5月份最高，8月份最低，5、 6月份是其产卵盛期．它的绝对怀卵数为18422 t 6033粒，每克 体重卵数为 501t 54粒乃体重，绝对怀卵数随体长与体重的增加 而增加．黑斑口虾结将卵产于洞穴内，产卵一般在傍晚，刚产出 一’．’ 的卵呈球形，卵粒之间由透明胶质物连接，排列成布状，再经过 附肢整理成簇状，挂于第5胸节腹面之前．巳抱卵的虾姑一般不 再出洞觅食．刚产出卵的卵径为 553675 u m，在正常水温范围内， 胚胎发育所需时间与水温成反比，在25刁3℃下，胚胎发育需6－ 12天． 5、黑斑口虾姑幼体在水温24．5－29℃下，经过27天的发育 二 A 变态，经历 3相（11期）的假蚤状幼体变成仔虾结．第 1相幼体体 表柔软，身体弯曲呈倒立状，不具游泳能力，多匍匐在水底或亲 体洞穴内，靠附肢作间歇运动，以卵黄为营养。头胸甲国呈近椭 圆形，中间刺强大，仅具第一、二胸肢与第一至四腹肢。第厂相 幼体能游泳与摄食，趋先上浮。头胸甲柔软呈梯形，第二触角刚 毛数增多，出现了第三至八对胸肢、第五至六腹肢及尾扇．第＊ 相幼体全身透明，趋先性减弱，活动于水体的中下部，游泳能力 很强，常在水中翻转或倒主；头胸甲及刺坚硬，手握有针刺感， 各附肢基本完善，腹部宽度增加，尾扇进一步发展。仔虾结身体 转为肉色，头胸甲两侧刺及中间刺消失，腹部明显增宽，且大于 头胸甲宽，尾扇发达，体形与生活习性与成体相似．随着幼体发 育，胃蛋白酶活力增加，在第7期出更多还原

【Abstract】 In this paper , the anatomy and histology of male and female reproductive systems , larvae development and technology of artificial reproduction of Oratosquilla kempi were conducted ,which was of great theory significance to the reproductive biology of Oratosquilla kempi and great practical significance to its mass production. The main results now showed as follows:1.During stage I of ovarian development, transparent ovarian lobes were sticky to gut without pigment, its ovogonium were in the reproductive cortices or the ovarian matrix nearby . At the beginning of stage II, the ovarian lobes began to develop and became grey, yellow and extended .At the same time the previtellogenic oocytes increased in volume with some vilelline. Ova can be seen clearly in ovarian lobes and became orange during stage III , the stage of vitellogenesis . The main character of stage IV was the increasing size of ovary and the separated mature ova. The results showed that O. kempi had a S-shaped proliferation district in the ovarian center generated from reproductive cortices of peripheral zone of ovary, which was different from other crustacean . The duration of ovogonium’s periodical development doesn’t end until the postvitellogenesis .This was the reason why O. kempi had longer spawning time than other crustacean2. The size of sexual cells,of nucleus and thenuclear-cytoplasmic ratio were different at different developing stages of ovary. Nucleus increased greatly at stage I and stage II, while from the beginning of stage III, the volume of sexual cells increased faster than nucleus. We may concluded that stage III was the key period forO. kempitoform and deposit nutritional material -vitellus , and the key points of culture parent O. kempi focused on early and middle stages of ovarian development,and that was the time from ovogonium to the forming of vitellus.3.The male reproductive system of O. kempi was consisted of V-shaped spermary ,deferensoid ,male gonopores , petasma and subsidiary gland. The appearance of spermary was V-shaped ,which was different from those H-shaped spermary of other crustacean .The histology studies indicated that there was no muscle cells in the wall of testis, so they were named as deferensoid. The spermatogenesis can be observed along the whole tubular testis. The spennatogonia , spermatocytes I , spermatocytes II and spermatozoa were arranged from basic membrane to cavity of testis. There was no spermatogonia in deferensoid and petasma. The size order of male reproductive cells at different stags was inverse to female reproductive cells. It can be deduced from the electronic microscope photos of sperm cells that what O. kempi ovulated was sperm cells not spermatozoa, the development of sperm cells to spermatozoa maybe happened in female O. kempi.4.The smallest biologiacal female and male examined had a total length of 8.3cm or 8.0 cm respectively. The mating season of 0. kempi was at the end of autumn or next spring above 15癈. The ovarianmature coefficient got highest in May and lowest in August , and the peak spawning period occurred in May-June. The absolute reproductive capacity was 18422 + 6033, and that was directly proportional to the total length .The eggs of O. kempi were laid in caves ,usually at evening. The eggs were connected each other just like a piece of cloth by transparent material and then changed into cluster and was hung on the abdominal head of O. kempi. The diameter of new spawned egg was 553-675 y m. The period of embryonic development is 6~12days under25~33癈 , and was inverse ratio to water temperature.5. The development of alima larva can be divided in 3 phases (or 11 stages ),and last 27d under24. 5-29癈. Stage I larva had first and second thoracic appendages and first to fourth pleopods , so it can’t swim but can sprawl intermittently by appendage in the caves, its carapace was soft and oval. The StageII larva had third-eighth thoracic appendage and fifth-sixth pleopod and tail fan, and can swim and eat . stage III larva are transparent a更多还原