The shoots grow 5-7 leaves from trees or cuttings, with a length of 5- 10 cm, and the shoots are removed. Rinse with tap water for 3-5 minutes, soak in 70% alcohol for 30-40 seconds, rinse with distilled water for 2-3 times, then put in 0.5% mercuric chloride solution for disinfection for 7- 10 minutes, and finally rinse with sterile water for 5-6 times.
Production and screening of culture medium
(1) shoot tip medium1/2ms medium 6- benzylpurine (6-BA) 0.5-lppm tea acetic acid (naa)0.0 1ppm hydrolyzed milk protein (lh) 100ppm sucrose 2% agar. Under the pressure of 98.06 kPa, autoclaved for 20 minutes, and cooled for later use. The stem tips of white feather, red banana, white friendship, riesling and other varieties were cultivated by Shandong Wine Grape Research Institute with this formula, and the survival rate was over 83.5%.
Square 2 1/2ms medium gibberellin (ga)0. 1ppm indoleacetic acid (1ba)0.59pm 6 nodal singing lppm kinetin (kt)0.5ppm adenine sulfate (ade)4ppm agar 0.6% sucrose 0.2% medium, bud mutation (kangtai).
(2) Stem and leaf differentiation medium 1 1/2ms medium gibberellin 1ppm indolebutyric acid 0.5ppm 6- nodosum 1ppm sucrose 2% agar 0.6%, and the hydrogen ion concentration was adjusted to 1000 nm/L (ph6.0). Sterilize under high pressure of 98.06 kPa for 20 minutes. The plant protection institute of Liaoning Academy of Agricultural Sciences has a survival rate of more than 90% for Peking University, Hongfutu, Heiaolin and Jufeng varieties.
Formula 2 adopts b5 macroelement medium, other elements in ms medium, indoleacetic acid (1aa) 0.01kloc-0/ppm 6-benzylpurine 0.02—0.59pm adenine (a) 2; Ppm or hydrolyzed milk protein 1000ppm sucrose 2%-3% agar 0.7%, hydrogen ion concentration 630.9- 1585 nm mol/L (pH 5.8-6.2), put in a triangular bottle, sealed with two layers of sulfuric acid paper, and the concentration of hydrogen ion is 98.06-168. The formula used by Henan Horticultural Research Institute has a good effect on the survival of early japonica varieties in Beijing.
(3) Young root differentiation medium 1 sterilized with bs macroelement medium sucrose 4% indoleacetic acid 1ppm 6- benzylpurine 0.08ppm agar 7% and hydrogen ion concentration 1000 nanomole/liter (ph6) for 20 minutes, and then cooled for later use. The Institute of Plant Protection of Liaoning Academy of Agricultural Sciences used this formula to insert single bud segments from shoot tips of Red Fuji, Heiaolin, Jufeng and other varieties into sterilized triangular flask culture medium. The bud level was 3.5 mm above the surface of the culture medium, and each flask was inserted with 1-3 stem segments, and then cultured in the dark for 48 hours. Then switch to intermittent illumination (illumination 10 hour per day under two 40-watt fluorescent lamps), and keep the temperature at 23-28℃. After 20 days, the average length of young roots was 99.0, 86.3 and 78.0 mm, respectively.
In Formula 2, sucrose 4% indoleacetic acid 0.4ppm 6- benzylpurine 0.06ppm agar 7% was used in 1/2ms medium, and the hydrogen ion concentration was 1000 nm/L (ph6). After autoclaving, inoculate Peking University, Gonggong Rabbit, Heiaolin and Chenfeng. After 20 days, the average length of young roots was 94.0 and 36.5 respectively. This culture medium has a good effect on reproduction of Peking University (Institute of Plant Protection, Liaoning Academy of Agricultural Sciences).
(3) Inoculation (in vitro culture of stem segments) on an ultra-clean workbench, cutting thoroughly sterilized spare buds into single-bud stem segments with 2 cm nuclei, and inserting the lower part into sterilized in vitro culture medium. The bud level is 3-5 mm above the surface of the culture medium, and 65,438+0-3 stem segments are inserted into each tube, which is cultured in the dark for 48 hours, and then switched to intermittent illumination (under two 40W fluorescent lamps, the illumination is 65,438/day).
Fourth, subculture: cut the test-tube seedlings into single bud segments with leaves without disinfection and insert them into human culture medium. Generally, it begins to take root in 10- 15 days and grows into 5.6-leaf seedlings in 30-35 days. Under suitable conditions, 1 year can breed tens of thousands of test-tube seedlings.
5. The test-tube seedlings are tempered and transplanted. After growing, they are moved to a greenhouse or a plastic stretcher and tempered under natural light. The temperature should be 20-25℃. After two weeks, remove the test tube plug and let it exercise in natural light, temperature and humidity for 4-5 days. Use tweezers to hold the petiole and take the seedlings out of the culture medium, wash off the residue of the culture medium, plant them directly in sandy loam or coarse sand in the small arch shed of greenhouse or greenhouse, and pour them through with bottom water. The humidity in the shed is kept at about 90%. The ground temperature at 10 cm should be stabilized at around 15℃, and the illumination should be 4000-5000 le. 15-20 days later, when the young leaves turn green, they can be transplanted into the field, and the survival rate is over 80%. If the temperature in the shed is too high, it is necessary to ventilate the reed curtain to cool down, spray cold water, and improve the air humidity and soil humidity in the shed, so as to maintain the suitable conditions for hardening seedlings and reasonable pruning.
Trim in time. Chucun Vineyard adopts three main vines with fan-shaped pruning (3 main vines and 6 ~ 7 side vines). Winter pruning is mainly based on short branch pruning, combining medium and short branches, and single branch regeneration.
The cutting length of the main vine was about1.2m. As a result, 2 ~ 3 buds were cut off from the mother branch, leaving a proper amount with a thickness exceeding 1cm. After the grapes germinate in spring, wipe the buds in time. When the buds grow to about 5 cm, remove the dense and useless new buds. When the inflorescence can be seen around 10 cm, remove the redundant development branches. When the height is 15 ~ 20cm, the branches are fixed, with an average of 8 ~ 10 new shoots per square meter, the spacing between new shoots is about 15 ~ 20cm, and the amount of shoots left per mu is about 8600. Two days before flowering, the stone removal of fruit branches was completed. Generally, when the new shoots above the inflorescence grow to 10 leaves, 6-8 leaves are left for coring, and only 1 secondary branches are left at the top of the fruiting branches, and all the others are smoothed. The remaining secondary buds are left for repeated coring until they stop growing and all the secondary secondary buds are removed. Results The mother shoots should be cut to death at one time, so that all nutrients can be supplied to the ear. After fruit setting, each branch leaves 10 ~ 1 1 leaf. There are generally no vegetative branches and extended branches. When it is really necessary to be used as a spare branch, the core can be picked repeatedly in 6 ~ 7 leaves, and the secondary branch can be picked repeatedly in 6 ~ 7 leaves to promote the maturity of branches and improve the quality of winter buds.
Sparse inflorescences and whole panicles
Adjust the load reasonably. The principle of ear-leaving is to leave 1 ear about 0.6 cm thick at the base of new shoots, 2 ears thick at 1 cm, no ear at weak branches, no ear below 30 cm from the ground, pinch off the ear tip 1/3 in time at the initial flowering stage and remove the secondary ear and tendrils to keep the ear at 500-600 g. When adjusting the load, it is necessary to
Rational use of fertilizer and water
Keep the tree strong. Apply decomposed organic fertilizer (mule manure, sheep manure and chicken manure) 1500 kg and calcium superphosphate 50 kg per mu every year. Topdressing 5 times a year, mainly nitrogen fertilizer after seedling release, mainly phosphorus and potassium fertilizer before flowering, mainly phosphorus and potassium fertilizer in small fruit stage, mainly phosphorus and potassium fertilizer in coloring stage. After harvest, phosphorus and potassium fertilizer are the main ingredients, with an appropriate amount of nitrogen fertilizer, each plant is 0.3 ~ 0.4 kg. In addition, foliar fertilizer is sprayed after flowering, once every half month 1 time, and 9 ~1time throughout the year. Nongjiabao and Lufeng 95 were mainly used in the early stage, and potassium dihydrogen phosphate was mainly sprayed in July. Watering depends on soil moisture. In case of drought in June and July, water should be replenished in time, and frozen water should be poured before wintering.
Strengthen pest control
Keep the blade intact. Spraying 5 Be stone sulfur mixture before burying and overwintering to reduce the source of pests and diseases in the next year, spraying 1 times the same concentration stone sulfur mixture before germination after grapes are unearthed; Spraying 1 ~ 2 times a year to control the occurrence of pests; The broad bean grain of the fruit is large enough for the fruit coloring period. Combined with foliar spraying fertilizer and mixed bactericide to prevent and control white rot and black pox. Spraying ammonium phosphate and metalaxyl at later stage to control downy mildew.