199 results about "Pollen tube" patented technology

The invention discloses a liquid medium for vitro scutellaria root pollen germination and a method for testing the activity of scutellaria root pollen. For the liquid medium for vitro scutellaria root germination, distilled water is used as a solvent, and the liquid medium comprises the components of BK, 100g/L cane sugar and 150g/L PEG4,000, the pH value of the liquid medium is 5.8. The tested results of pollen activity is stable and reliable by utilizing the liquid medium for vitro scutellaria root pollen germination and a pollen tube growth testing technology, the ratio of the pollen germination reflects the proportion of pollens with vitality, and the observation and the measurement of the pollen tube reflect the advantages and disadvantages of physiologic conditions of pollen grains, thus the technology for testing vitro scutellaria root pollen germination and pollen tube growth provides a reliable and effective method for the measurement of the activity of the scutellaria root pollen germination.

The invention discloses a plant expression vector and application thereof. The plant expression vector comprises an expression box expressing a tfdA gene, wherein the expression box comprises a CaMV35S promoter, a tfdA gene and a terminator that are sequentially connected; the nucleotide sequence of the tdfA gene is a sequence 2 in a sequence table, and the nucleotide sequence of the CaMV35S promoter is a sequence 1 in the sequence table. The construction of the vector of the invention has very important application values on the construction of the gene recombinant expression vector of the cotton, the gene transformation (especially providing incontestable evidence for the transgenes of pollen tubes in China), later generation screening of the transgenes and the whole cotton gene engineering for the research of the cotton transgenes.

The invention relates to a fluorescence microscopy method for rapidly and efficiently observing a camellia plant pollen tube. The method comprises the following steps: (1) dissecting and separating a pistil material of a camellia plant to be observed into an operation unit; and (2) sequentially soaking the separated material in NaClO solution and NaOH solution until the material is transparent and gets softened, dyeing through a water-soluble aniline blue dye solution, flaking by employing a double-glass slide attachment both-sided observation method, and finally performing pollen tube fluorescence microscopy. The method is easy, convenient and feasible to operate, the experiment efficiency is greatly improved, and the observation effect is optimized.

This invention relates to a method for increasing the conversion efficiency of pollen tube pathway method by using agrobacterium tumefaciens virulence gene. The method comprises: combining or mixing agrobacterium tumefaciens virulence gene and the target gene to be converted (green fluorescent protein gene) in vitro, packaging with liposome, and converting plants, especially cotton by using pollen tube pathway method. The method combines agrobacterium-liposome-pollen tube pathway technique, and has obviously increased protoplast conversion efficiency. Plasmids are not only protected by liposome when entering embryo sacs, but also encapsulated by the toxic protein to be expressed when entering egg cells, which can improve the stability of exogenous DAN in cell conversion process, realize precise cutting in T-DNA region, and increase the conversion efficiency.

The invention relates to chemical male sterilization, pollen-tube pathway and pollen-mediated transformation technologies, and belongs to the technical field of molecular breeding and gene engineering. The method comprises the following steps of: performing chemical male sterilization treatment on a crop T0 of a budding stage; cutting stigmas of buds; mixing fertile pollen by using the pollen-tube pathway technology, namely using target gene-containing agrobacterium-mediated infection solution (added with 5 percent of sucrose and 500muL/L of SilwetL-77) with OD600 of 0.8; and dipping an absorbent cotton ball in the bacterial solution, coating the bacterial solution to the stigma cut ovaries, performing shading treatment for three days to realize normal growth, and finally collecting selfing seeds. Herbicide resistance screening is performed on the generation T1, polymerase chain reaction (PCR) detection is performed on resistant plants, and positive plants are determined. The method has the characteristics of low cost, simplicity in operation, no need of expensive instruments and equipment, high transformation rate (1 to 5 percent) and the like, is suitable for any flowering plant, and can be applied to large-scale crop genetic transformation.

The invention provides a method for quickly identifying cross-compatibility of rose hybrida. The method is characterized by comprising the following steps of: collecting and storing pollen, performing castration, performing artificial pollination, cutting a sample to be identified, fixing the cut sample in FAA stationary liquid, transferring the cut sample to a 70 percent ethanol solution after 24 hours, taking out for washing, placing in an 8mol/L NaOH solution, performing water bath at the temperature of 55 to 60 DEGC for 15 to 20 minutes, and then standing at the temperature of 20 to 28 DEGC for 3 to 3.5 hours, soaking into water for at least 1 hour, transferring to a 0.1 percent dye solution of aniline blue, dyeing for 2 to 3 hours, observing stigma callose deposition, pollen germination and growth of pollen tubes by using a fluorescence microscope to identify the cross-compatibility of rose hybrida. By the method, the cross-compatibility of rose hybrida can be identified within 30 to 48 hours; and moreover, the method is simple and easy to operate and can effectively reduce the waste of breeding resources and improve breeding efficiency.

The invention relates to a culture medium for improving germination and growth of Kumaiti Armeniaca vulgaris Lam pollen. The culture medium is prepared by adding gibberellin, naphthylacetic acid, indoleacetic acid or 2,4-dichlorphenoxyacetic acid serving as a growth regulator into agar, sucrose and boric acid which serve as basic culture medium components, or adding potassium nitrate, magnesium sulfate, urea, calcium sulfate, ammonium molybdate, sodium molybdate, ferric sulfate, zinc sulfate or copper sulfate serving as a chemical element into agar, sucrose and boric acid which serve as basic culture medium components in a mass ratio. The culture medium solves the problems of low germination rate of the pollen and slow growth of pollen tubes, and an effective path for improving the germination rate is found for Kumaiti. Through the culture medium, the germination and the growth of Luntai Armeniaca vulgaris Lam pollen are improved; and the culture medium is easy and convenient to operate, low in cost, short in period and high in germination rate, and has wide application prospect in drupe tree biology.

The invention discloses a plant pollen tube cell calcium ion ultrastructure positioning method which mainly comprises the following steps: (1) preparing a quantitative immobilizing liquid and washing liquid; (2) immobilizing and washing a material; (3) immobilizing and washing secondarily; (4) dehydrating and transiting; (5) carrying out resin permeating, embedding and polymerizing; (6) directly observing by using an electron microscope without dyeing the material. Due to the adoption of the method, the problems that cell turgor pressure needs to be adjusted, abnormal reaction can be caused between potassium pyroantimonate and buffer liquid, calcium pyroantimonate precipitation displacement or dilution can be caused by material immobilization and aftertreatment, Ca<2+> precipitate observation can be affected because of inappropriate dyeing, and the like can be solved, and organelle can be clearly distinguished without dyeing completely.

The invention discloses a labeling method of an apple pollen tube microfilament. The labeling method comprises the following steps: cultivating apple pollen, curing the apple pollen, performing enzyme treatment on the apple pollen, treating the apple pollen with a surfactant, performing dye labeling, and performing scanning and imaging. The labeling method has the beneficial effect that the labeling method of gymnosperm pollen tube microfilament is referred and improved on the basis of the labeling method of pear pollen tube microfilament, dyes enter cells easier through curing, enzymolysis of cell walls and the surfactant, the interference of cell outer wall autofluorescence due to insufficient enzymolysis as TRITC-Phalloidin is applied are avoided when FITC-Phalloidin labeling is applied, so that relatively complete apple pollen tube microfilament can be stored; moreover, the labeling method is efficient and quick, can be used for obtaining relatively clear and bright pictures, thus filling up the blank of the apple pollen tube microfilament labeling, providing reference for labeling microfilament in rosaceae and angiosperm pollen tube, and achieving a high application value.

The invention discloses dragon fruit pollen suspending liquid and a preparation method and a pollination method thereof. The dragon fruit pollen suspending liquid disclosed by the invention can not only provide nutrients needed by in vitro pollen by containing sufficient nutritional agents, but also promote the pollen to germinate and grow in a short time by being blended with a suitable plant growth regulator, is stable in property and can be placed for a long time without generating a lamination phenomenon. The dragon fruit pollen suspending liquid disclosed by the invention is easy and convenient to use and improves the pollination efficiency by 2-3 times by adopting a spraying pollination method which is simple and easy to operate. The pollination method of the dragon fruit pollen suspending liquid can effectively promote the germination of the pollen and the growth of a pollen tube and achieve the pollen germination rate of more than 34.5%, is favorable to the pollination and fruit setting of dragon fruits and achieves the fruit setting rate of more than 96.5%.

The invention discloses a method for extracting nucleuses of a pear pollen tube. The method comprises the following steps: 1) collecting pollen, that is, collecting anther in a big bud stage and drying the anther; 2) culturing the pollen; 3) releasing nucleuses, that is, grinding the pollen with a single-cell screen and carrying out filtration by using a dual-cell screen so as to release the nucleuses; 4) purifying the nucleuses by using Percoll density gradient centrifugation; and 5) rinsing Percoll. The method does not relate to enzyme extraction of protoplast or conventional technologies of grinding and filtering, and has the advantages of simple and fast operation, low cost, an obvious purification effect, etc.; the purified nucleuses are hardly contaminated by cytoplasm, can not onlybe used for preparation of DNA fibers but also be used for proteomics research of nucleuses, and have important significance to research on growth, metabolism and self-incompatible response of pear pollen tubes.

The breeding process of high temperature resisting cotton hybrid belongs to the field of crop hybrid breeding technology. The breeding process includes the following steps: 1. selecting parents for hybridizing to obtain cotton hybrid F1; 2. planting cotton hybrid F1 based on the randomized block experiment design and field surveying single plant cotton boll number and boll forming rate; 3. indoor in vitro pollen culture to measure the pollen germination rate and pollen tube growth speed; and 4. screening through principal component analysis. The said process can culture high temperature resisting cotton hybrid with high yield, powerful adaptability and high cotton quality.

The invention discloses a tobacco pollen medium as well as a preparation method and application thereof. The preparation method of the tobacco pollen medium comprises the following steps of: picking flowers of a tobacco male parent from a bud containing period to an initial blooming period; removing anthers, placing on an indoor ventilating position, and cooling for 24 hours; collecting stigmas; drying under the condition of 30 DEG C; grinding, and then screening through a sieve of 60 meshes so as to obtain the tobacco pollen medium. The tobacco pollen medium and pure pollen are uniformly mixed according to the weight ratio of 1:2 and then used for the artificial pollination of tobaccos. The tobacco pollen medium can promote the germination of the pollen and the extension of pollen tubes, enhance the successful ratio of pollination, increase the output of tobacco seeds in unit area, reduce the usage amount of the pollen and effectively reduce the production cost of the tobacco seeds; and in addition, the invention changes wastes into valuables and has the advantages of easiness and convenience for operation, low cost and good application prospect.

The invention provides a breeding method capable of improving the content of potassium in tobacco leaves. According to the invention, total DNA of the high potassium plant pokeweed is introduced into Honghuadajinyuan, a cultivated variety of tobacco, by using a pollen tube pathway method, which results in substantial increase of the content of potassium in later-generation tobacco leaves of Honghuadajinyuan, and therefore, a rich source of variation is provided for breeding and improvement of new varieties of tobacco and innovation of germplasm resources.

The invention discloses a fluorescence labeling method for free calcium ions of a cotton pollen tube, and belongs to the technical field of biology. The fluorescence labeling method comprises the following steps: perforating glass slide to make a small chamber for culturing the pollen tube; culturing pollen in the small chamber, and sprouting the pollen tube; loading Fluo-3AM into the cultured pollen tube by a low temperature incubation method; and observing dynamic variation of Ca2+ in the pollen tube under a confocal laser scanning microscopy. The invention establishes a method for researching the concentration change of the Ca2+ in the cotton pollen tube by using the confocal laser microtechnique, has simple operation and good labeling effect, plays an important role in the growth of the cotton pollen tube and fertilization reproduction research, and has important practical application value.

The invention discloses a lactuca sativa L. pollen in vitro germination culture medium and a method for measuring pollen activity. The lactuca sativa L. pollen in vitro germination culture medium uses distilled water as a solvent and comprises components of ME3 (Monnior), PEG4000 (Polyethylene Glycol) and saccharose. The method comprises the following steps: (1), collecting pollen, namely, selecting an inflorescence flowering in that day, transferring into a freshness protection package filled with clear water, and bringing back in a room; (2), preparing a pollen in vitro liquid culture medium; (3), spreading and culturing pollen, namely, horizontally placing a glass slide loaded with the liquid culture medium on an experiment platform, removing petals of a capitulum by using tweezers, stripping off a synandrium stamen and then taking an anther out, enabling the anther to slide on the surface of the culture medium to ensure that pollen is uniformly spread and distributed on the surface of the culture medium in a single layer and a pollen tube grows; (4), measuring the pollen activity, namely, after in vitro culturing, observing, and counting a pollen germination rate, wherein the pollen germination rate reflects a proportion of vital pollens. The method is rapid, simple and easy to operate, good in culturing effect, and large in practical application value; abundant vital germinating pollens are obtained, and the pollen activity is effectively measured.

The invention discloses a sorghum variety breeding and cultivating method. The method includes the following steps of introducing a non-toxic broad-spectrum anti-disease gene PYH157 and an insect-resistant gene CpTI into sorghum through a pollen tube channel method, selecting a positive plant, obtaining seeds of the plant, sun-drying, soaking and stirring the seeds, sowing the seeds, applying farmyard manure and base fertilizer, conducting early thinning and uniformizing to keep strong seedlings, conducting dispersing to keep the seedlings not dense but loose, pulling away sick, weak and hybrid seedlings, finally singling 2-3 seedlings in each pot according to the soil fertility, fertilizer applying level and sowing density when 5-6 leaves grow on each seedling, conducting hole-applied fertilizer dressing after the final singling, conducting the first time of intertillage soil loosening while weeding, conducting deep elongation and booting fertilizer dressing before elongation, and conducting the third time of intertillage ridging. The disease resistance of sorghum crops is improved, the growth cycle of sorghum is shortened, the maturity speed is higher, the disease resistance of the plant is high, yield is increased, quality is better, pods are plump, and plant diseases and insect pets are remarkably reduced.

The invention discloses a composition of plant direct gene transformation, transforming method and appliance, which is characterized by the following: making the component include goal gene and free nucleic acid component of adjusted and controlled element, transforming buffer liquid and transforming assistance component; co-culturing; soaking; surface-daubing; injecting; dripping; spraying; transforming with dip flower method or through genetic gun bombardment, supersonic wave and electric shock method. This component can be used to receptor plant cell direct transformation of fertilized ovum, suspended cell, protoplast, single-layer cell and diachyma cell and can be used to seed, receptor plant pollen, pistil, flower pillar, pollen pipe and ovary, which possesses impotent meaning for plant genetic seeding, variety improvement and gene functional analysis.