43 results about "Female Gametophytes" patented technology

The invention discloses a method for cloning and culturing seaweed gametophytes, which comprises the following steps of: cutting seed seaweed pieces with mature sporangia; using sterilized absorbent cotton to dip the boiled seawater for cleaning the seed seaweed pieces; respectively soaking the pretreated seed seaweed pieces with sterilized double-resistant seawater for 30 minutes, sterilized seawater solution with the potassium iodide concentration being 1.5% for 10 minutes, and sterilized seawater for 20 minutes; carrying out free spore diffusion and filtration in a serum bottle; and pouring the filtered spore water into a vertical staining jar which is provided with a glass slide, after the free spores are attached to the glass slide to form embryo spores, pouring the spore water out, filling sterilized double-resistant seawater, using a micropipette to separate single male and female gametophytes to the serum bottle, then using sterilized double-resistant seawater to culture, and thus obtaining the unicellular seaweed gametophyte clone group. The seaweed gametophyte which is cultured by the method has stable clone characteristics and high survival rate, can be effectively prevented from being contaminated, and is suitable for long-term storage.

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 and sea tangle and enhance the quality of the hybrid undaria pinnatifida and sea tangle.

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.

It is intended to provide a novel unialgal culture strain showing a high culture efficiency of a large-sized red alga which is immature, can be stored and cultured over a long period of time and has at least one of the following properties, i.e., producing a physiologically active substance at a high yield, showing a high growth speed of the alga body, and being highly capable of absorbing nutritional salts. Namely, an immature unialgal culture strain originating in a large-sized red marine alga which is characterized by showing no matured female gametophyte in nature but showing matured tetrasporophyte alone and grows in a natural marine water area containing fresh water. This unialgal culture strain is constructed by collecting the matured sporophyte, cutting the sporophyte and allowing to stand to thereby release spores, culturing the released spores and continuously proliferating and culturing even after the growth of an upright body from the germinated spore.

The invention discloses a kelp gametophyte solid phase culturing storing method. It adopts sea belt male and female gametophyte clone, and includes the following steps: preparing solid phase culturing culture medium; smashing gametophyte clone; inoculating gametophyte; solid phase culturing; solid phase storing and resuscitation enlarging foster. The invention can make liquid culture storing long, anaphase operation simple, and provides the possibility of applying spaceflight breeding technique in marine alga breeding.

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 for cultivating undaria pinnatifida and using the undaria pinnatifida to treat red tide. The method is characterized in that according to a sign before the occurrence of the red tides, that is, the temperature is increased by 2 DEG C within 7 days, a method different from a biological restraint method is adopted; before the outbreak of the red tides, the cultivatedundaria pinnatifida is added into the sea to absorb nutritional components in the sea water to alleviate the eutrophication of the sea water, and thus the occurrence of the red tides is avoided. The method includes: performing induced cultivation on female gametophyte to obtain young sporophyte, and carrying out aerated feeding cultivation to obtain mature sporophyte; releasing the sporophyll of the sporophyte into the sea to obtain zoospores, and placing seedling curtains attached with the zoospores in fresh sea water; when the germinated zoospores develop into female gametophyte, co-cultivating male gametophyte and the female gametophyte attached to the seedling curtains to complete the development and fertilization process; transferring the young sporophyte to a relevant sea area for cultivation after the young sporophyte reaches 50-100 micrometers to obtain the undaria pinnatifida and treat the red tides.

The invention provides a novel method for breeding kelp seedlings with a gametophyte cloning method, comprising the following steps of: respectively separating a female cell on a female gametophyte and a male cell on a male gametophyte under a microscope by using a capillary tube separation technology; then respectively cloning the female cell and the male cell by using a cloning vegetative propagation technology to respectively grow the female cell and the male cell into female and male gametophytes; continually breeding the gametophytes; when the gametophytes are bred to a fixed amount, damping a formed cell cluster by a cell stamping machine; mixing the female and male gametophytes and respectively releasing ova and sperms by the female and male gametophytes to spontaneously fertilize;putting fertilized zygotes into nutritive salt to be cultured; and after 10 weeks, obtaining seedlings with the height of about 1.5 cm. According to the invention, a fertilization process and the control of temperature and illumination are realized by manual operation, so that the growth period of offspring seeds is greatly shortened; meanwhile, the female cell and the male cell are separated from the gametophytes by using the capillary tube separation technology to clone and culture, so that the quality and the yield of the seedlings are improved, the cultivation of the kelp can be satisfied; and by utilizing the novel method, the cost for breeding the kelp seedlings is low.

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 relates to the field of economical brown kelp seaweed varietal complexity, in particular to a method for detecting varietal complexity in kelp summer seedling refrigeration water process. The method comprises the steps that a seedling rope which is inoculated with a female gametophyte clone is put in a culture medium or a kelp seedling refrigeration water circulation system to be cultured, parthenogernesis sporophore is obtained when the seedling rope is put in the culture medium, and offspring sporophore is obtained when the seedling rope is put in the kelp seedling refrigeration water circulation system; morphological characteristics and genetype of the offspring sporophore on the seedling rope in the refrigeration water circulation system are detected and compared with those of the parthenogernesis sporophore so as to determine whether varieties are hybrid or not in the seedling process. According to the detection method, it is verified for the first time that in the kelp summer seedling production process, due to the fact that low temperature circulation water is used in common, the varietal complexity is definitely caused.

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 relates to genetic breeding, in particular to a method for constructing a permanent F2 sporophyte population in undaria pinnatifida, and further provides a method for obtaining a undaria pinnatifida permanent F2 sporophyte population by utilizing full-sib F2 gametophyte clone pairwise hybridization. The method comprises the following steps: respectively separating male and female gametophyte clones (F1 gametophyte) from homozygous parent sporophytes, hybridizing the obtained male and female gametophyte clones to generate F1 sporophytes, selecting a strain of released zoospores after the F1 sporophytes are mature, and performing separating to obtain a large number of F2 gametophyte clones; and randomly selecting female gametophyte clones and male gametophyte clones with the same number from the F2 gametophyte clones, and then completing hybridization of the F2 male gametophyte clones and the F2 male gametophyte clones on the randomly selected gametophytes in a female-male random one-to-one pairing mode, so as to obtain a permanent F2 sporophyte population. The permanent F2 population constructed by the invention has important significance in development of basic genetic research, new variety cultivation and the like in important economic brown seaweed undaria pinnatifida.

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 facilitating ovulation of kelp female gametophytes. The method includes the steps: resuscitating kelp germplasm filaments in seawater, crushing kelp germplasms, andfiltering crushed liquid to obtain filament fragments with the lengths of 3-6 cells; performing aerated suspension culture on the filament fragments under the conditions of the illumination intensityof 6000+ / -300lx, the illumination cycle of L:D=12h:12h and the temperature of 15+ / -1 DEG C to collect eggs; culturing the collected eggs to obtain young kelp sporophytes, culturing the young kelp sporophytes, and performing culture by running water at the stage of young sporophytes of two rows of cells. According to the method, female kelp germplasms can ovulate in 11-13 days, ovulation time is shortened, ovulation rate is high, ovulation synchronization is good, ovulation rate within two days can reach 65-70%, and egg development rate can reach 70-80%. According to the built culture method of young sporophytes, the aberration rate of the young sporophytes can be reduced to 50-60%.

The invention discloses a method for increasing the proportion of drynaria roosii female gametophytes. The method comprises the steps that after drynaria roosii spores are sown aseptically, then placed under an incandescent lamp to be cultured to form protonemata, placed under LED blue light or LED red light or LED white light or LED green light at a plate stage, a heart-shaped prothallium formation stage or a sexual organ differentiation stage respectively to be cultured and placed under the incandescent lamp at the other stages. Compared with other LED light qualities, a large proportion (80%) of unisexual female gametophytes are generated when the LED blue light is preferentially selected for treatment in the heart-shaped prothallium formation stage, and the other 20% gametophytes are asexual, so that the selfing behavior in the breeding process is avoided, and the method is the quite ideal light quality treatment mode for obtaining the unisexual female gametophytes.

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 discloses a kelp gametophyte DNA solution preparation method and a sex determination method. The preparation method comprises the steps that kelp gametophyte is added with sterile deionized water, full grinding is conducted, after boiling water bath is conducted, room-temperature standing is conducted, supernatant is transferred into a sterile centrifuge tube, by means of a female specific primer HFM4 and a male specific primer Msj68-16-3, a round of double-primer PCR amplification is conducted, and the sex and confusing situation of multiple varieties or strains of kelp gametophyte are identified rapidly and accurately. The method of preparing a kelp gametophyte DNA solution easily and rapidly without using any molecular reagents is provided, a kelp female gametophyte specific marker is developed, the pertinence is high, the amplification efficiency is high, and the stability is good; by combining a kelp male gametophyte specific marker developed by a lab, the method ofidentifying the sex by means of the kelp gametophyte DNA solution is built, the sex of the kelp gametophyte can be detected, and whether or not the male and female mixed phenomenon occurs is determined.

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 discloses a method for promoting rapid development of gametophytes of Czostera crassipes, and particularly relates to a method for promoting rapid development of gametophytes of Czostera crassipes by adopting blue light as a light source, adopting CO (NH2) 2 as a nitrogen source and adding 0.5 mg / L of FeC6H5O7-Fe into a culture solution in the development stage of the gametophytes, and adopting the optimal development conditions of the gametophytes of Czostera crassipes under other conditions. The method provided by the invention can effectively accelerate the development process of the gametophytes of the chlamydomonas palustris, promote the synchronous development of the male and female gametophytes and improve the sporophyte conversion rate, thereby improving the utilization rate of the gametophytes, reducing the time required for seedling culture, improving the stock-out specification of seedlings and reducing the cost of seedling breeding.

The invention relates to genetic breeding, in particular to a method for constructing a double haploid (DH) sporophyte population in undaria pinnatifida. The invention still further relates to the method for obtaining an undaria pinnatifida DH sporophyte population by utilizing parthenogenesis and hermaphrodite gametophyte selfing. The method comprises the following steps of selecting female gametophytes with parthenogenesis capability for cloning and male gametophytes capable of generating a hermaphroditism phenomenon for cloning, hybridizing the female gametophytes and the male gametophytes to generate F1 sporophytes, selecting a strain to release zoospores after the F1 sporophytes are mature, and separating to obtain a large number of F2 gametophytes for cloning; and selecting the female gametophytes for cloning, obtaining a female DH sporophyte strain by inducing parthenogenesis, selecting the male gametophytes for cloning, and obtaining a male DH sporophyte strain by inducing hermaphroditic phenomenon and selfing, and thereby obtaining the DH sporophyte population. The DH population constructed by the method has important significance in developing basic genetics research, new variety cultivation and the like in important economic brown algae undaria pinnatifida.

The invention discloses a saccharina japonica female gametophyte specific molecular marker FSMSJ-1294 and application thereof, and belongs to the field of genetic engineering. A sequence of the saccharina japonica female gametophyte specific molecular marker FSMSJ-1294 is shown as SEQ ID NO: 5, sequences of amplification primers of the saccharina japonica female gametophyte specific molecular marker FSMSJ-1294 are shown as SEQ ID NO: 3 and SEQ ID NO: 4, and the saccharina japonica female gametophyte specific molecular marker FSMSJ-1294 can be used for identifying a parental origin of a parthenogenetic saccharina japonica sporophyte and the gender of offspring of the parthenogenetic saccharina japonica sporophyte. A cell line cloned by female and male gametophytes of saccharina japonica is cultured, gametophyte cells are collected, genome DNA of the gametophyte cells is extracted, PCR amplification is carried out by using the primers to obtain an amplification product, and after electrophoresis detection, if a specific band with the length of 1294bp can be detected, the cell line is determined as a saccharina japonica female gametophyte cell line. The saccharina japonica female gametophyte specific molecular marker FSMSJ-1294 is stable in identification result, rapid and convenient to operate, high in stability, and low in detection cost.

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 relates to a method for improving the induction rate of embryogenic calluses of pinus koraiensis, in particular to the method for improving the induction rate of the embryogenic calluses of the pinus koraiensis. The invention aims to solve the problem of low induction rate of the embryogenic calluses of the pinus koraiensis at present. The method comprises the following steps of pretreating pinus koraiensis cones, and culturing the pinus koraiensis cones at 4 to 6 DEG C for 28 to 35 days to obtain cultured pinus koraiensis cones; treating the cultured pinus koraiensis cones to obtain sterilized female gametophytes, then inoculating the sterilized female gametophytes into a callus induction culture medium, and performing dark culture at the temperature of 21 to 25 DEG C for 30 to 60 days to obtain cultured embryogenic calluses; mixing the cultured embryogenic calluses with a basic culture medium, oscillating and centrifuging, and filtering to obtain a dispersed embryogenic cell cluster; and placing the dispersed embryogenic cell cluster on a mature culture medium, and performing dark culture for 40 to 60 days to obtain a somatic embryo. The method for improving the induction rate of the embryogenic calluses of the pinus koraiensis can be obtained by the invention.

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.