技术领域technical field
本发明涉及一种海洋生物的养植方法，具体涉及一种红壳色品系马氏珠母贝的制种方法。The invention relates to a method for cultivating marine organisms, in particular to a seed production method for a red-shell color strain of Pinctada martensii.
背景技术Background technique
研究表明，个体在生长过程中对贝壳颜色的选择与其生长环境(包括水域的温度、盐度等)、地理位置和生理因素都有着一定的相关性。根据对一些贝类的研究报道，壳色多态性是简单的孟德尔式遗传，但是壳色的选择与生长性状、生理性状和一些经济性状是否具有必然的联系，都没有明确的结论。Studies have shown that there is a certain correlation between the individual's choice of shell color in the growth process and its growth environment (including the temperature and salinity of the water area, etc.), geographical location and physiological factors. According to some shellfish research reports, shell color polymorphism is a simple Mendelian inheritance, but there is no clear conclusion whether the selection of shell color is necessarily related to growth traits, physiological traits and some economic traits.
马氏珠母贝(Pinctada martensii Dunker)隶属于双壳纲、珍珠贝科、珠母贝属，是一种重要的海水珍珠养殖贝类，其壳表呈淡黄色，常有数条黑褐色放射纹。其它壳表颜色较少见。在马氏珠母贝的遗传育种研究过程中，我们发现一个家系在壳色上表现出较大差异，壳表颜色为红色，具有较明显的红色放射纹，珍珠层外缘呈红色，而且该性状能遗传。Pinctada martensii Dunker (Pinctada martensii Dunker) belongs to Bivalvia, Pearl Oysteraceae, and Pinctada. Other case colors are less common. During the research on the genetics and breeding of Pinctada martensii, we found that a family showed a large difference in shell color, the shell surface color was red, with obvious red radial patterns, the outer edge of the nacre was red, and the Traits can be inherited.
日本马氏珠母贝Pinctada fucata martensii的壳色包括红色、褐色、黄色和棕色，Wada K.T.等(1983，1990，1994，1996)发现在野生和养殖群体里白色贝非常稀少，曾在一些自交群体里发现过白色个体。白壳色性状有可能是由隐性基因控制的，而且其产生的珠不含黄色素，将白色个体作为小片贝，对于培育优珠是有利的，但白色壳近交系的个体总重和成活率远低于白壳/棕色壳杂交系的个体总重和成活率。海湾扇贝(Argopecten irradians Lamarck)的壳色能够稳定的遗传，受环境因素影响较小(Kraeuter J.et al，1984；Elek J.A.et al，1985，1990；Adamkewicz L.et al，1988)，理论上通过定向选育可以培育出不同壳色的海湾扇贝新品种。Gary等(1980)对贻贝Mytilus edulis不同壳色群体进行杂交，形成的分离家系的研究中，发现两个群体间的壳色多态性存在相当频率的差异，并且在不同群体间棕色个体比蓝色个体要小10-20％。但John等(2004)对太平洋牡蛎的研究发现，其平均个体重、成活率及生长率性状都与壳色素及外套膜缘色素没有相关性。The shell colors of Pinctada fucata martensii include red, brown, yellow and brown. Wada K.T. et al. (1983, 1990, 1994, 1996) found that white shellfish were very rare in wild and cultured populations, and had been used in some self-bred White individuals have been found in the colony. The trait of white shell color may be controlled by a recessive gene, and the pearls produced by it do not contain yellow pigment. It is beneficial to use white individuals as small shellfish for breeding high-quality pearls, but the total weight of white shell inbred lines and The survival rate was much lower than the total individual weight and survival rate of the white/brown shell hybrids. The shell color of the bay scallop (Argopecten irradians Lamarck) can be inherited stably and is less affected by environmental factors (Kraeuter J. et al, 1984; Elek J.A. et al, 1985, 1990; Adamkewicz L. et al, 1988), theoretically New varieties of bay scallops with different shell colors can be bred through directional selection. Gary et al. (1980) crossed different shell color populations of the mussel Mytilus edulis to form a segregation family. They found that there were considerable frequency differences in the shell color polymorphism between the two populations, and the ratio of brown individuals between different populations Blue individuals are 10-20% smaller. However, John et al. (2004) found in the study of Pacific oysters that the average individual body weight, survival rate and growth rate traits were not correlated with shell pigments and mantle margin pigments.
我国在贝类壳色研究方面，已根据不同壳色群体的生长状况，对海湾扇贝和皱纹盘鲍数量性状进行定向选育，获得了表型明确、性状优良的新品系。Zhang等(2003，2005)以壳色为标记，目前已经培育出纯橘红色、纯黑色、纯紫色、纯白色等不同颜色的海湾扇贝，建立了海湾扇贝“壳色-数量性状复合选择和自交-定向选育-小群体平衡”的育种模式，培育出“中科红海湾扇贝”新品种，95％以上的子代个体为桔红壳色，成活率提高15％-20％，生长速度提高10％-15％，出肉率增加10％以上，壳厚也明显增加。在养成阶段，相同日龄的不同壳色间，白色家系个体显著的大于橙色和紫色个体；高温的7、8月份，白色家系的存活率最高(由于白色贝壳吸收光热比橙色紫色个体吸收的少，更容易适应高温强光的环境)，并发现在幼虫阶段，海湾扇贝个体的生长性状和存活率与壳色之间不存在主要联系，可能是因为在幼虫阶段个体都具有相同的遗传基础，而且还是透明的，不具备分泌色素到壳的功能。刘晓等(2003)采用杂交育种的方式已经育成了皱纹盘鲍“中国红”品系，壳色基因已达纯合，建立了12个“中国红”同胞或半同胞家系，并进行了规模繁育，为皱纹盘鲍的养殖产业提供了一个很好的发展方向。目前，尚未见红壳色品系马氏珠母贝制种方法的报道。In terms of research on shell color of shellfish in my country, according to the growth status of different shell color populations, the quantitative traits of bay scallop and wrinkled plate abalone have been selected, and new strains with clear phenotype and excellent traits have been obtained. Zhang et al. (2003, 2005) have bred bay scallops of different colors, such as pure orange red, pure black, pure purple, pure white, etc., using shell color as a marker, and established the "shell color-quantitative traits compound selection and automatic selection method" of bay scallops. The breeding mode of "cross-directed breeding-small group balance" has bred a new variety of "Zhongke Honghai Bay Scallop". More than 95% of the progeny individuals are orange-red shell color, the survival rate is increased by 15%-20%, and the growth rate is fast. Increase by 10%-15%, the meat output rate increased by more than 10%, and the shell thickness also increased significantly. In the growth stage, among different shell colors at the same age, the individuals of the white family are significantly larger than those of the orange and purple individuals; in July and August when the temperature is high, the survival rate of the white family is the highest (because the white shell absorbs more light and heat than the orange and purple individuals) It is easier to adapt to the environment of high temperature and strong light), and found that in the larval stage, there is no main relationship between the growth traits and survival rate of bay scallop individuals and shell color, which may be because individuals in the larval stage have the same genetic basis , but also transparent, does not have the function of secreting pigment into the shell. Liu Xiao et al. (2003) have bred the "China Red" strain of wrinkled plate abalone by means of cross breeding, the shell color gene has reached homozygosity, established 12 "China Red" compatriots or half-sib families, and conducted large-scale breeding , providing a good development direction for the aquaculture industry of wrinkled plate abalone. At present, there is no report on the seed production method of the red shell color strain Pinctada martensii.
发明的内容content of the invention
本发明的目的是提供一种具有明显表型性状——红壳色品系马氏珠母贝的制种方法，该品系贝可作为细胞小片贝或育珠贝，以培育出不同于通常的白色或黄色珍珠颜色，使珍珠颜色多元化，提高红壳色的纯度，最高可达96％，不需通过转基因等复杂技术导入外源基因，因此该产品对生物和环境安全没有影响。The purpose of the present invention is to provide a seed production method with obvious phenotypic traits—the red-shelled strain of Pinctada martensii, which can be used as cell lamellae or pearl oysters to breed different white species. Or yellow pearl color, so that the pearl color is diversified, and the purity of the red shell color can be improved, up to 96%. It does not need to introduce foreign genes through complex technologies such as genetic modification, so this product has no impact on biological and environmental safety.
本发明的的目的是这样实现的：一种红壳色品系马氏珠母贝的制种方法，它依次包括下列步骤：(1)制种：通过单交培育具有红壳色的种贝，即从已培育出的高纯度红壳色的家系(或从野生或养殖群体内)选出红壳色特征明显的雌雄贝各1，进行单交，对各单交组分别培育；(2)扩繁：以上述方式培育几代，其红壳比例达到90％以上时，开始扩繁生产种苗：各单交组配对进行群体交配，或单交组的雌与野生或养殖群体中的雄群交。The purpose of the present invention is achieved like this: a kind of seed production method of red-shell color strain Pinctada martensii, it comprises the following steps successively: (1) seed production: by single-cross cultivation with the seed shellfish of red-shell color, That is, select 1 male and female shellfish with obvious red-shell color characteristics from the family of high-purity red-shell color that has been cultivated (or from wild or cultured groups), perform single-crossing, and cultivate each single-crossing group separately; (2) Expansion: Cultivate several generations in the above-mentioned way, and when the proportion of red shells reaches more than 90%, start to multiply and produce seedlings: each single-cross group is paired for group mating, or the female of the single-cross group and the male in the wild or cultured group group sex.
扩繁生产的种苗作为细胞小片贝或育珠贝，以培育出红壳色珍珠。The seedlings produced by the expansion are used as cell shellfish or pearl oysters to cultivate red-shell color pearls.
所述(1)中的制种，依次包括下列步骤：The seed production in (1) comprises the following steps successively:
a.亲贝培育a. Broodling
从一个红壳色家系(从野生或养殖群体内)中挑选红壳特征明显、性腺较饱满个体，贝龄1.5-2龄；于室内蓄养3-5天，当水温较低时，升温至25℃培养，当水温达25℃以上时，进行常温培养；蓄养期间，每天换水一次，且投喂充足的饵料，促进性腺的进一步发育成生理成熟状态；Select individuals with obvious red shell characteristics and full gonads from a red shell color family (from wild or cultured groups), and the shellfish age is 1.5-2 years old; keep them indoors for 3-5 days, when the water temperature is low, raise the temperature to 25 Cultivate in ℃, when the water temperature reaches above 25℃, culture at room temperature; during the breeding period, change the water once a day, and feed enough bait to promote the further development of the gonads into a physiologically mature state;
b.制取法收集精卵和授精b. Preparation method to collect sperm and eggs and insemination
授精采用单交，分别吸取精卵于盛有过滤海水的烧杯中，收集的精卵经筛绢过滤，除去杂质及组织碎片，并迅速将卵加入盛有0.005-0.007％的氨海水的平底玻璃容器中，停留10分钟，然后加入适量的精子，并混匀让其受精；授精发育水温在20℃-28℃之间；换水两次以洗去多余的精子；The insemination adopts single crossing, and the sperm and eggs are respectively drawn into the beaker filled with filtered seawater. The collected sperm and eggs are filtered through a sieve to remove impurities and tissue fragments, and the eggs are quickly added to the flat-bottomed glass filled with 0.005-0.007% ammonia seawater Stay in the container for 10 minutes, then add an appropriate amount of sperm, and mix well to fertilize; the water temperature for insemination and development is between 20°C and 28°C; change the water twice to wash away excess sperm;
c.幼虫收集及培养c. Larva collection and culture
待幼虫大量上浮后，用虹吸法将上浮幼虫收集到育苗池；水体中幼虫的数量控制在2～8个/ml；前三天采用加水的方法，以后用套有一定网孔大小筛绢的换水器进行换水，换水量为育苗池总水体的1/3-1/2，每天至少换水一次；上下午各投饵一次，早期幼虫投喂金藻；进入壳顶初期，加投扁藻；变态附着后，饵料以扁藻为主；培养时连续充气，换水时暂停充气；After a large number of larvae float up, use the siphon method to collect the floating larvae into the nursery pond; the number of larvae in the water body is controlled at 2 to 8/ml; the method of adding water is used for the first three days, and the sieve with a certain mesh size is used later. The water changer is used to change the water, the water change is 1/3-1/2 of the total water body of the nursery pond, and the water is changed at least once a day; the feed is fed once in the morning and afternoon, and the early larvae are fed with golden algae; Flat algae; after abnormal attachment, the bait is mainly flat algae; continuous aeration during cultivation, and suspension of aeration when changing water;
d.采苗d. Picking seedlings
当用显微镜观察有20～30％的幼虫出现色素点时，投放片状采苗器或网状采苗器；采苗器附着有大量幼虫而育苗池水体中还有大量幼虫浮动时，把附着幼苗的采苗器转移到另一育苗池进行培养，而在原来的育苗池中投放采苗器进行第二次采苗；当所有的幼虫附着后(约需十几天)，换水时即取去换水器上的筛绢；When 20-30% of the larvae have pigmented spots under the microscope, put in a sheet seedling picker or a net seedling picker; The seedling picker of the seedlings is transferred to another seedling pond for cultivation, and the seedling picker is put into the original seedling pond for the second seedling picking; Remove the sieve from the water changer;
e.收苗、海上养殖e. Harvest seedlings, sea culture
当幼苗个体长到1.5-2.0mm时，片状采苗器上的幼苗用海绵或手轻轻地擦洗下来，然后将适量幼苗放入一级笼中(约60目)，出池下海；而网状采苗器，直接装笼出海；海上养殖按常规进行，需及时分笼，检查壳色纯度。When the individual seedlings grow to 1.5-2.0 mm, the seedlings on the sheet-shaped seedling picker are gently scrubbed off with a sponge or hands, and then an appropriate amount of seedlings are put into a first-level cage (about 60 mesh), and they are taken out of the pool and into the sea; Shaped seedling picking device, directly put into cages and go out to sea; marine culture is carried out according to routine, and it is necessary to divide cages in time and check the purity of shell color.
扩繁生产的种苗作为细胞小片贝或育珠贝育珠，育珠指用该培育出的贝为细胞小片贝来插核育珠。其术前处理、插核技术、休养及珍珠养成与常规方法相同。基本方法为：The seedlings produced by the expansion are used as small cell shellfish or pearl shellfish to cultivate pearls, and pearl cultivation refers to using the cultivated shellfish as cell shellfish to insert nucleus and grow pearls. The preoperative treatment, nucleus insertion technique, recuperation and pearl cultivation are the same as the conventional method. The basic method is:
1、母贝的术前处理1. Preoperative treatment of mother shellfish
采用密集低吊的方法，抑制生殖腺的发育及降低其生理机能Intensive low-hanging methods are used to inhibit the development of gonads and reduce their physiological functions
2、制作外套膜小片2. Making small pieces of mantle
一般选用1龄左右的生长良好的贝作小片贝，取出左右外套膜的下方游离部分，放置在玻璃板上，并切成小方块形。Generally, well-grown shellfish of about 1 year old are selected as small pieces of shellfish, and the lower free parts of the left and right mantles are taken out, placed on a glass plate, and cut into small cubes.
3、插核3. Insert core
在母贝的足基部用开口刀在表皮上开一切口，把珠核和小片送入切口部，使小片的外上皮向珠核。Make an incision on the epidermis with an open knife at the foot base of the mother oyster, and send the bead nucleus and small pieces into the incision, so that the outer epithelium of the small pieces faces the bead nucleus.
4、休养4. Rest
手术后的母贝吊养在休养场，时间为一个月左右。After the operation, the female oysters are suspended and raised in the rest field for about a month.
5、育珠和收珠5. Pearl cultivation and collection
休养期过后，把育珠贝转入养殖海区进行常规的管养，一般需养殖12-18个月后开贝收珠。After the recuperation period, the pearl oysters are transferred to the culture sea area for conventional management. Generally, it takes 12-18 months to open the shells and collect the pearls.
本发明的优点和积极效果在于：Advantage and positive effect of the present invention are:
通过本发明可较简单地培育出红壳色的品系，并提高红壳色的纯度，最高可达96％，不需通过转基因等复杂技术导入外源基因，因此该产品对生物和环境安全没有影响。Through the present invention, the red-shelled strain can be bred relatively simply, and the purity of the red-shelled color can be increased up to 96%. It is not necessary to introduce exogenous genes through complicated techniques such as transgenics, so the product has no biological and environmental safety. Influence.
具体的实施方式specific implementation
通过实施例进一步说明了本发明，但不构成对本发明的限制。The present invention is further illustrated by examples, but not construed as limiting the present invention.
实施例1.Example 1.
(1)制种(1) Seed production
2.亲贝培育2. Breeding of broilers
从一个具有红壳色的家系(养殖群体)中挑选红壳特征明显、性腺较饱满个体，贝龄2龄；于室内蓄养5天，水温达25℃以上，进行常温培养。蓄养期间，每天换水一次，且投喂充足的饵料，促进性腺的进一步发育成生理成熟状态。From a family (breeding group) with red shell color, select individuals with obvious red shell characteristics and full gonads, and the shellfish age is 2 years old; they are kept indoors for 5 days, and the water temperature is above 25°C, and they are cultured at room temperature. During the breeding period, the water is changed once a day, and sufficient bait is fed to promote the further development of the gonads into a physiologically mature state.
精卵收集及授精Sperm and egg collection and insemination
精卵的收集采用剖取法。授精采用单交，分别吸取精卵于盛有过滤海水的烧杯中，收集的精卵经筛绢过滤，除去杂质及组织碎片。并迅速将卵加入盛有0.005％的氨海水的平底玻璃容器中，停留10分钟，然后加入适量的精子，并混匀让其受精。授精发育水温在26℃。换水两次以洗去多余的精子。Sperm and eggs were collected by dissection. The insemination adopts single crossing, and the sperm and eggs are respectively drawn into a beaker filled with filtered sea water. The collected sperm and eggs are filtered through a sieve to remove impurities and tissue fragments. And quickly add the eggs into a flat-bottomed glass container filled with 0.005% ammonia seawater, stay for 10 minutes, then add an appropriate amount of sperm, and mix well to allow them to be fertilized. The water temperature for insemination and development was 26°C. Change the water twice to wash off excess sperm.
幼虫收集及培养Larva collection and culture
待幼虫大量上浮后，用虹吸法将上浮幼虫收集到育苗池。水体中幼虫的数量控制在5-6个/ml。前三天采用加水的方法，以后用套有一定网孔大小筛绢的换水器进行换水，换水量为育苗池总水体的1/3，每天至少换水一次。上下午各投饵一次，早期幼虫主要投喂金藻；进入壳顶初期，加投扁藻；变态附着后，饵料以扁藻为主。幼虫培养时连续充气，换水时暂停充气。在培养过程中，应每天取一些幼虫在显微镜下检查幼虫的摄食及生长情况，以及时掌握饵料的投喂量。After a large number of larvae float up, the siphon method is used to collect the floating larvae into the nursery pond. The number of larvae in the water body is controlled at 5-6/ml. The method of adding water is used in the first three days, and then the water is changed with a water changer with a certain mesh size sieve. The water change is 1/3 of the total water body of the seedling pond, and the water is changed at least once a day. Feed once in the morning and in the afternoon, and the early larvae are mainly fed with golden algae; when they enter the shell top stage, they are fed with flat algae; after metamorphosis and attachment, the bait is mainly flat algae. Continuously inflate during larvae culture, and suspend aeration when changing water. During the cultivation process, some larvae should be taken every day to check the feeding and growth of the larvae under the microscope, so as to grasp the feeding amount of the bait in time.
采苗Picking seedlings
当有25％的幼虫出现色素点时(通过显微镜观察)，投放片状采苗器。(采苗器主要有两种，一种为片状采苗器，即用塑料片串成，有红色和黑色两种，黑色较好；另一种为网状采苗器，即为各种规格的胶丝网片、灯光网、旧鱼网等。)When 25% of the larvae had pigment spots (observed through a microscope), the sheet seedling picker was put in. (There are mainly two kinds of seedling picking devices, one is a sheet seedling picking device, which is made of plastic sheets, and there are two kinds of red and black, black is better; the other is a mesh seedling picking device, which is a variety of Standard glue wire mesh, lighting net, old fishing net, etc.)
采苗器投放后，随时观察幼虫附着动态，采苗器附着有大量幼虫而育苗池水体中还有大量幼虫浮动时把已附着幼苗的采苗器转移到另一育苗池进行培养，而在原来的育苗池中投放采苗器进行第二次采苗。所有的幼虫附着后(约需十几天)，换水时即取去换水器上的筛绢。After the seedling picker is put in, observe the larva attachment dynamics at any time. When there are a large number of larvae attached to the seedling picker and there are still a large number of larvae floating in the water body of the seedling pond, transfer the seedling picker that has attached seedlings to another seedling pond for cultivation. Put the seedling picker in the seedling pond for the second seedling picking. After all the larvae are attached (it takes about ten days), remove the sieve silk from the water changer when changing the water.
收苗、海上养殖Harvest Seedlings, Mariculture
幼苗个体长到大约1.5-2.0mm，出池下海。片状采苗器上的幼苗用海绵或手轻轻地擦洗下来，然后将适量幼苗放入一级笼中(约60目)。(采用网状采苗器时，一般直接装笼出海。)海上养殖按常规进行，需及时分笼，检查壳色纯度。The individual seedlings grow to about 1.5-2.0 mm, and go out of the pond and into the sea. The seedlings on the sheet seedling picker are gently scrubbed with a sponge or hands, and then put an appropriate amount of seedlings into a first-level cage (about 60 mesh). (When using a net-shaped seedling picker, it is generally directly put into a cage to go out to sea.) Mariculture is carried out as usual, and it is necessary to divide the cages in time and check the purity of the shell color.
实施例2.Example 2.
地点：深圳大亚湾，时间：2004年10月，以实施例1方式培育几代，其红壳比例达到90％以上时，开始扩繁生产种苗：各单交组配对进行群体交配，取具有红壳雌贝4只，雄贝4只，剖取法收集精卵，用0.006％的氨海水处理卵10分钟，然后加入精子在水温23℃下授精，而后用过滤海水两次换水。常规养殖，结果红壳比例80％。Place: Daya Bay, Shenzhen, time: October 2004, several generations were cultivated in the manner of Example 1, and when the red shell ratio reached more than 90%, the multiplication and production of seedlings began: each single-breeding group was paired for group mating, and the red shell was taken. There were 4 female shellfish and 4 male shellfish. Sperm eggs were collected by dissection, and the eggs were treated with 0.006% ammonia seawater for 10 minutes, then sperm were added to inseminate at a water temperature of 23°C, and then the water was changed twice with filtered seawater. Conventional breeding results in a red shell ratio of 80%.
实施例3.Example 3.
地点：深圳大亚湾，时间：2005年4月，与实施例2类似，取具有红壳雌贝1只，雄贝1只，剖取法收集精卵，用0.006％的氨海水处理卵10分钟，然后加入精子在水温24℃下授精，而后用过滤海水两次换水。常规养殖，结果红壳比例96％。Location: Daya Bay, Shenzhen, time: April 2005, similar to Example 2, take 1 female shellfish and 1 male shellfish with red shell, collect sperm eggs by dissection, treat the eggs with 0.006% ammonia seawater for 10 minutes, and then Add sperm to inseminate at a water temperature of 24°C, and then change the water twice with filtered seawater. Conventional breeding results in a red shell ratio of 96%.
实施例4Example 4
地点：深圳大亚湾，时间：2005年4月，与实施例2类似，取具有红壳雌贝1只，雄贝1只，剖取法收集精卵，用0.006％的氨海水处理卵10分钟，然后加入精子在水温24℃下授精，而后用过滤海水两次换水。常规养殖，结果红壳比例76％。Location: Daya Bay, Shenzhen, time: April 2005, similar to Example 2, take 1 female shellfish and 1 male shellfish with red shell, collect sperm eggs by dissection, treat the eggs with 0.006% ammonia seawater for 10 minutes, and then Add sperm to inseminate at a water temperature of 24°C, and then change the water twice with filtered seawater. Conventional breeding results in a red shell ratio of 76%.
实施例5.Example 5.
地点：深圳大亚湾，时间：2005年4月，取具有红壳雌贝1只，雄贝1只(非红壳色)，剖取法收集精卵，用0.006％的氨海水处理卵10分钟，然后加入精子在水温24℃下授精，而后用过滤海水两次换水。常规养殖，结果红壳比例89％。Location: Daya Bay, Shenzhen. Time: April 2005. One female shellfish and one male shellfish (non-redshell color) were taken, and sperm eggs were collected by dissection. The eggs were treated with 0.006% ammonia seawater for 10 minutes, and then Add sperm to inseminate at a water temperature of 24°C, and then change the water twice with filtered seawater. Conventional breeding results in a red shell ratio of 89%.
| Application Number | Priority Date | Filing Date | Title |
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| CNB2006100349573ACN100362912C (en) | 2006-04-12 | 2006-04-12 | A kind of seed production method of red shell color strain Pinctada martensii |
| Application Number | Priority Date | Filing Date | Title |
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| CNB2006100349573ACN100362912C (en) | 2006-04-12 | 2006-04-12 | A kind of seed production method of red shell color strain Pinctada martensii |
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| US6251438B1 (en)* | 1995-12-28 | 2001-06-26 | Centre National De La Recherche Scientifique (Cnrs) | Method of preparing active substances from nacre, products obtained which can be used in particular as medicaments |
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| US6251438B1 (en)* | 1995-12-28 | 2001-06-26 | Centre National De La Recherche Scientifique (Cnrs) | Method of preparing active substances from nacre, products obtained which can be used in particular as medicaments |
| US5807554A (en)* | 1997-04-11 | 1998-09-15 | Yng-Wong; Quing Non | Herbal formulations with nacre |
| CN1372790A (en)* | 2002-03-22 | 2002-10-09 | 中国科学院南海海洋研究所 | Artificially cultured triploid Hepu nacre and its pearl culturing method |
| US7062940B2 (en)* | 2002-12-13 | 2006-06-20 | Chi Huynh | Carved pearl |
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| CN1826880A (en) | 2006-09-06 |
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