43 results about "Female Gametophytes" patented technology

The invention relates to an undariapinnatifida cultivation method, in particular to an undariapinnatifida seedling cultivation method through parthenogenesis. The method comprises the steps that induction culture is conducted on female gametophytes until juvenile sporophytes reach more than 1mm, aerated feeding culture is conducted on the female gametophytes, then the life history of parthenogenesis sporophytes is completed, and mature sporophytes are obtained; sporophyls of the sporophytes are placed into sea water to be released to obtain planospores, after the color of the sea water where the planospores are obtained through releasing is changed into yellowish-brown, the planospores are attached to a seedling curtain, and then the seedling curtain is put into fresh sea water; after all the germinal planospores develop into the female gametophytes and are sufficiently grown, light intensity is lowered, the development of the gametophytes is delayed to enable the gametophytes to pull through the high-water-temperature summer; the male gametophytes and the female gametophytes attached to the seedling curtain are simultaneously cultured, and the development and fertilization processes are completed; after reaching about 200 micrometers, the juvenile sporophytes are moved to sea to be cultivated, and then undariapinnatifida cultivation is achieved. According to the undariapinnatifida seedling cultivation method through parthenogenesis, female gametophyte clonal update and enlarged cultivation can be achieved through the parthenogernesis life history, and it is of great significance in clonal preservation and crossbreeding.

The invention belongs to the technology field of the gene engineering, in particular relates to a specificity molecule marker which can distinguish laminaria female and male gametophyte. The specificity molecule marker is obtained through the following method: two pairs of primers of SEQ ID No1and SEQ ID No2 are designed according to 5'end flanking sequence of the laminaria female and female gametophyte gene groups, polymerase chain reaction amplification is performed to the deoxyribonucleic acid of the laminaria female and female gametophyte gene groups, and the deoxyribonucleic acid of specificity amplification fragment are reclaimed; large intestine rod competent strain JM109 is cloned through pMD19-T vector to obtain a laminaria female gametophyte specificity molecule marker Rf-763 and a laminaria male gametophyte specificity molecule marker Rm-239, and the sequences thereof are SEQ ID No3 and SEQ ID No4. The genders of the laminaria gametophyte, parthenogenesis laminaria sporophyte parental source and the genders of the offspring thereof can identified through utilizing the specificity molecule marker, and the laminaria gender differentiation in basis of the cell and molecule level can be studied.

The invention belongs to the technology field of the gene engineering, in particular relates to a specificity molecule marker which can distinguish laminaria female and male gametophyte. The specificity molecule marker is obtained through the following method: two pairs of primers of SEQ ID No1and SEQ ID No2 are designed according to 5'end flanking sequence of the laminaria female and female gametophyte gene groups, polymerase chain reaction amplification is performed to the deoxyribonucleic acid of the laminaria female and female gametophyte gene groups, and the deoxyribonucleic acid of specificity amplification fragment are reclaimed; large intestine rod competent strain JM109 is cloned through pMD19-T vector to obtain a laminaria female gametophyte specificity molecule marker Rf-763 anda laminaria male gametophyte specificity molecule marker Rm-239, and the sequences thereof are SEQ ID No3 and SEQ ID No4. The genders of the laminaria gametophyte, parthenogenesis laminaria sporophyte parental source and the genders of the offspring thereof can identified through utilizing the specificity molecule marker, and the laminaria gender differentiation in basis of the cell and moleculelevel can be studied.

The invention discloses an algae breeding method based on intergeneric hybridization, which is characterized by using temperature, illumination and other physical conditions and chemical elements in culture solution, such as nitrogen, phosphorus, iron and the like to treat algae gametophyte cells of different genera of Laminariales; synchronously developing different genera of female gametophytes and male gametophytes to form intergeneric heterozygotes; and developing the intergeneric heterozygotes to large sporophytes. The method has the advantages that great differences of forms, yields and economic components (algin, polysaccharides from Dunaliella salina) of different genera of algaes are utilized to culture artificial algae with the comprehensive advantages of two genera of algaes through intergeneric hybridization, thus obtaining novel algae varieties or strains having the advantage of comprehensive economic characters, and improving economic attributes of yield, economic components and the like.

The invention belongs to the field of seaweed molecular genetics, and relates to an enteromorpha gender specific molecular marker and application thereof. The enteromorpha gender specific molecular marker is characterized in that one enteromorpha female gametophyte DNA specific molecular marker and one enteromorpha male gametophyte DNA specific molecular marker are provided, wherein the nucleotidesequences of the enteromorpha female gametophyte DNA specific molecular marker and the enteromorpha male gametophyte DNA specific molecular marker are respectively SEQ ID NO. 3 and SEQ ID NO. 4, theSEQ ID NO. 3 is obtained by amplifying a primer sequence SEQ ID NO. 1, and the SEQ ID NO. 4 is obtained by amplifying a primer sequence SEQ ID NO. 2. The specific molecular marker is completed by detecting the specific fragment through PCR (polymerase chain reaction), and the process comprises the following steps of extracting to-be-detected enteromorpha genomic DNA, performing PCR amplification,and performing agarose gel electrophoresis detection on an amplification product. The specific primer is obtained by referring to existing high-throughput data result in a laboratory, combining genomeDNA extraction, target fragment amplification and recovery, cloning by using pGEM-T vector and escherichia coli competent strain TOP10 and the like. By means of the specific molecular marker, ploidyof a large number of enteromorpha algae and gender of gametophytes can be rapidly, simply and accurately identified, and an important molecular tool is provided for enteromorpha life history researchand variety breeding.

The invention provides a method capable of improving masson pine somatic embryo inductivity. The method comprises the work procedures: pretreating explant, sterilizing the expalnt and inducing a somatic embryo; collecting masson pine superior tree immaturate cones as the explant, pretreating and sterilizing the explant, peeling female gametophyte in the explant cones in a sterile mode, flatly putting the female gametophyte in an induction medium and putting into a suitable environment to be cultured, wherein the immaturate cones are healthy and have no pest and disease damage, zygotic embryo of the immaturate cones are in a cleavage polyembryony period, and an embryogenic callus induction culture period is 3 to 4 weeks; finally, obtaining a masson pine embryonic cell line which has quickness in growth and exuberant vigor. According to the method disclosed by the invention, a series of optimized explants are treated and sterilized, then female gametophyte in the cones is peeled and induced to form a lot of embryonic cell lines with exuberant vigor, powerful material resources and technical supports are established for further developing masson pine somatic embryo seedling culture, germplasm innovation and bioengineering seed breeding, and better economic benefits, social benefits and ecological benefits are achieved.

The invention belongs to the technology field of gene engineering, in particular relates to a sequence characterized amplified regions molecule marker which can distinguish laminaria female gametophyte. The sequence of the molecule marker is SEQ ID No.4. The molecule marker is obtained through the following method: a primer is designed according to an earth wire Y chromosome conservative region and polymerase chain reaction amplification is performed to the laminaria female and female gametophyte gene groups; the original primer is extended to be a novel primer SEQ ID No.3 according to the sequencing result of the amplification product; a sequence characterized amplified regions molecule marker F-494 of the laminaria female gametophyte is obtained through utilizing the primer to perform polymerase chain reaction amplification to the laminaria female and female gametophyte gene groups and the sequence thereof is SEQ ID No.4. The gender of the laminaria gametophyte can be identified rapidly and efficiently through utilizing the sequence characterized amplified regions molecule marker, and the parthenogenesis laminaria sporophyte parental source and the gender of the offspring thereof can be identified.

The invention discloses an artificial insemination method for Filicinae, and belongs to the technical field of plant artificial insemination. In order to solve the problem of low seedling rate of pteridophytes, the invention provides an artificial insemination method of Filicinae, which comprises the following steps: inoculating spores into a culture medium, obtaining female gametophytes and malegametophytes of the Filicinae by using a conventional spore culture method, after a sexual organ is mature, sucking mature sperms and a prepared induction liquid, sequentially dropwise adding the mature sperms and the prepared induction liquid to a mature archegonium of the female gametophyte, and enabling the sperms to enter the archegonium to be combined with ova under the guidance of the induction liquid, so as to complete artificial insemination. According to the sperm taking and artificial directional insemination technology provided by the invention, the fertilization rate of the gametophyte can reach 30%-50% and is 3-5 times that under conventional culture conditions, the phenomenon that a single gametophyte has two embryonic developments often occurs, and the hybridization technology of pteridophytes is simpler, more convenient and easier to implement.

The invention provides a method for improving the induction rate of somatic embryos of Pine massoniana, which comprises the steps of explant pretreatment, explant disinfection and somatic embryo induction, and collects healthy Pine massoniana with no pests and diseases and zygotic embryos in the stage of cleavage and multiple embryos. The immature cones of the tree are used as explants. After the explants are pretreated and disinfected, the female gametophytes in the explant cones are aseptically stripped, placed flat on the induction medium and placed in a suitable environment Medium culture, embryogenic callus induction culture period: 3-4 weeks, and finally obtain the embryogenic cell line of Pinus massoniana with rapid growth and vigorous vitality. The present invention undergoes a series of optimized explant treatment and disinfection, and then strips the female gametophytes in the cones for induction, forming a large number of vigorous embryogenic cell lines, which is for further development of Pinus massoniana body embryo seedling breeding, germplasm innovation and biological Engineering breeding has established strong material resources and technical support, and has good economic, social and ecological benefits.

The invention relates to an undariapinnatifida cultivation method, in particular to an undariapinnatifida seedling cultivation method through parthenogenesis. The method comprises the steps that induction culture is conducted on female gametophytes until juvenile sporophytes reach more than 1mm, aerated feeding culture is conducted on the female gametophytes, then the life history of parthenogenesis sporophytes is completed, and mature sporophytes are obtained; sporophyls of the sporophytes are placed into sea water to be released to obtain planospores, after the color of the sea water where the planospores are obtained through releasing is changed into yellowish-brown, the planospores are attached to a seedling curtain, and then the seedling curtain is put into fresh sea water; after all the germinal planospores develop into the female gametophytes and are sufficiently grown, light intensity is lowered, the development of the gametophytes is delayed to enable the gametophytes to pull through the high-water-temperature summer; the male gametophytes and the female gametophytes attached to the seedling curtain are simultaneously cultured, and the development and fertilization processes are completed; after reaching about 200 micrometers, the juvenile sporophytes are moved to sea to be cultivated, and then undariapinnatifida cultivation is achieved. According to the undariapinnatifida seedling cultivation method through parthenogenesis, female gametophyte clonal update and enlarged cultivation can be achieved through the parthenogernesis life history, and it is of great significance in clonal preservation and crossbreeding.

The invention relates to a method adopting undaria pinnatifida female gametophytes saved and cultured indoors and sea tangle female and male gametophytes to be mutually used as male parents and female parents and cutting the gametophytes. The method comprises the following steps of: mixing according to the mode of the undaria pinnatifida female gametophytes * the sea tangle male gametophytes and the sea tangle female gametophytes * undaria pinnatifida male gametophytes; cutting into algae segments of 200 micrometers by a tissue pounding machine, and then scattering on a vinylon seeding bed; and cultivating the vinylon seeding bed at temperature suitable for the growth and the maturity of the undaria pinnatifida and sea tangle gametophytes, wherein after culturing for 7 days to 10 days, the sea tangle female gametophytes are mature to form oocysts and ovulate ova which are respectively fertilized with sperms released by mature sea tangle and undaria pinnatifida male gametophytes so as to germinate hybrid seedlings. The invention can sufficiently utilize the heteroses of the distant hybridization of undaria pinnatifida and sea tangle; in addition, obtained hybrids have inheritable characteristics of the undaria pinnatifida and the sea tangle, high stress resistance, fast growth and good quality and can greatly increase the cultivation output of the hybrid undaria pinnatifida andsea tangle and enhance the quality of the hybrid undaria pinnatifida and sea tangle.

The invention relates to a method for crossbreeding of undaria pinnatifida, in particular to a method for crossbreeding of the undaria pinnatifida by using unisexual planospores. Female gametophytes are subjected to parthenogernesis induction to generate sporophores, meanwhile, hermaphroditic gametophytes are subjected to development cultivation to generate selfing sporophores, when the sporophores are longer than 1mm, the sporophores are changed into inflating cultivation conditions and cultivated to sexual maturity. Mature sporophyls are placed in sea water for releasing the planospores, the planospores are attached on a seeding curtain after the color of sea water turns into tawny, and the seeding curtain is then placed in fresh sea water. The planospores released by the sporophores generated by parthenogernesis are female planospores, and the planospores released by the hermaphroditic gametophytes are male planospores. According to the method, the male and female gametophyte hybridization technique is improved by the male and female planospore hybridization technique, and the method has great significance in applying monoclonal crossbreeding to the production practice in large scales.

The invention belongs to the field of seaweed molecular genetics, and in particular relates to a method for identifying the sex of kelp gametophytes. It is characterized in that: Laminaria kelp female gametophyte and male gametophyte specific molecular markers are respectively named SJF and SJM, and their nucleotide sequences are respectively SEQ ID NO: 5 and SEQ ID NO: 6. SEQ ID NO: 5 is amplified from the primer Sf sequence of SEQ ID NO: 3; SEQ ID NO: 6 is amplified from the primer Sm sequence of SEQ ID NO: 4. The invention is accomplished by detecting the above-mentioned specific fragment; the steps include extraction of the Laminaria gametophyte genome DNA to be detected, PCR amplification, and agarose gel electrophoresis detection of the amplification product. In summary, the present invention can clearly identify whether the Laminaria gametophyte is pure female, pure male or mixed sex, which makes up for the deficiencies of the prior art.

The invention discloses a method for preventing and controlling benthic diatom pollution in the production of wakame gametophyte seedlings. In the invention, the male and female gametophytes before the pond in the production of wakame gametophyte seedlings are pre-developed and broken, and then they are attached to the pond, so that the wakame gametophyte and juvenile sporophyte compete with the benthic diatoms on the seedling curtain. Occupy an advantage to prevent benthic diatom pollution in the production of wakame gamete seedlings. The technology of the invention has good effect of preventing and controlling benthic diatoms, is convenient and applicable, and can also shorten the cultivation time of the seedlings in the nursery pond and save the production cost.

The present invention relates to genetic breeding, specifically a method for constructing doubled haploid (DH) sporophyte populations in Undaria pinnatifida, and further to obtain Undaria pinnatae by using parthenogenesis and selfing of hermaphrodite gametophytes Methods for DH sporophyte populations. Select female gametophyte clones with parthenogenetic ability and male gametophyte clones that can produce hermaphroditic phenomena, and cross the two to produce F 1 sporophyte, after it matures, select a strain to release zoospores, and obtain a large amount of F 2 Gametophyte clone; select the female gametophyte clone among them, obtain the female DH sporophyte strain by inducing the occurrence of parthenogenesis, select the male gametophyte clone among them, and obtain the male DH sporophyte strain by inducing the occurrence of hermaphroditism and selfing, Thereby obtaining DH sporophyte population. The DH population constructed by the invention is of great significance for basic genetics research and new variety cultivation in important economical brown algae Undaria pinnatae.