CN1053345A

CN1053345A - Isolated rapid reproduction method of wintersweet

Info

Publication number: CN1053345A
Application number: CN 90104477
Authority: CN
Inventors: 傅萼辉; 徐惠珠
Original assignee: Wuhan Institute of Botany of CAS
Current assignee: Wuhan Botanical Garden of CAS
Priority date: 1990-06-15
Filing date: 1990-06-15
Publication date: 1991-07-31
Also published as: CN1018328B

Abstract

A kind ofly make explant with the elite stand axillalry bud of growing up, adopt the minimal medium and the additional plant hormone of autogamy to make up to the stripped rapid breeding method of plum blossom, its advantage is: grow seedlings to seedling to flowering time short (2-3), plant has that strong stress resistance, life-span are long, the position of blooming is good, small product size is little, is easy to sell, and is beneficial to commercialization production.

Description

Isolated rapid reproduction method of wintersweet

Invention relates to the Woody flower method for quickly breeding, does not relate to plant variety.People exsomatize to plum blossom with additional plant hormone combination by a kind of minimal medium and breed fast.

In the prior art, do not see the stripped rapid breeding method of plum blossom, find no the report of pass through inventor's retrieval yet.Only have minority kinds such as Chinese rose, cuckoo to realize quick breeding in Woody flower, Prunus (PRUNUS) does not all have to realize breeding fast though plant has nearly 30 kinds to pass through obtaining regeneration plant from in-vitro culture.From growing seedlings to seedling to flowering time long (6-7), plant resistance is poor under field conditions (factors) in existing conventional plum blossom grafting, and the life-span is short, the position height of blooming, and unfavorable moulding, small product size is big, is difficult for selling.

The object of the present invention is to provide a kind of improved method for quickly breeding, having shortened plum blossom grows seedlings to seedling to flowering time, be the improved seeds of China tradition famous flower-plum blossom and the batch production production of precious wild type, and the high-grade export-oriented commodity plum blossom that forms potted plant (or potted landscape) has been opened up approach.

The invention is characterized in that adopt the autogamy medium of low inorganic salt concentration, plant hormone is combined as compound, explant adopts the elite stand axillalry bud of growing up, vegetative progeny good reliability.

In the procedure that the present invention proposes, adopt the peripheral wounded in the battle flower bud branch of the elite stand that grows up, be cut into 15-20 centimetre length plugged ear, handle, insert in the greenhouse casting bed routinely without plant hormone, treat that bud is sprouted and then adopt axillalry bud and make explant, under aseptic condition, strike off outer scale with pocket knife, the alcohol disinfecting with 70% 3 minutes, again with 0.1% mercuric chloride solution sterilization 5 minutes, inoculate with behind the aseptic water washing 4 times.With the medium for preparing with 100 milliliters of conical flask packing (every bottled 40-50 milliliter), through high temperature (120-125 ℃), high pressure (1.1KG/CM ²) sterilize to cool off and then on superclean bench, inoculate.Every combination 3-5 bottle, every bottle of 5 explants.After the inoculation culture placed the quiet cultivation of culturing room.Culturing room's temperature is controlled at (22-28 ℃), uses the daylight lamp as light source, and intensity of illumination is 1000-2000LUX, illumination every day 12 hours.Just cultivate, the auximone combination of the phytocytomine of the additional high concentration of minimal medium and low concentration induces simple bud to grow up to the bud of growing thickly; Bud carries out successive transfer culture to growing thickly, the sprout tuber that then will grow thickly is transferred to successive transfer culture on minimal medium and the additional plant hormone, every bottle of culture can grow the sturdy no offspring of 4-6 strain, to no offspring root induction, continue to cultivate with the medium of autogamy and additional plant hormone, every strain grows the sturdy root of 2-6 bar from stem after 25 days, when little strain root length arrives 1-1.5 centimetre, just from medium, take out, after running water cleans, transplant in the basin soil, put the greenhouse and support seedling, move on to outdoor management when treating plant length again to 15-20 centimetre.

The present invention is the method for breeding fast, grows seedlings to seedling to lack (2-3) to flowering time, and plant has strong stress resistance, and the life-span is long, and the position of blooming is good, is convenient to moulding, and small product size is little, is easy to sell, and is beneficial to commercialization production.

Fig. 1: the bud of growing thickly of axillalry bud differentiation

Fig. 2: the seedling of successive transfer culture

Fig. 3: induce the seedling seedling of taking root

Fig. 4: 2 years living test tube plantlet sprays of plum blossom

Embodiment:

Adopt the autogamy medium of low inorganic salt concentration to form: ammonium nitrate (NH by following composition ₄NO ₃) 1200-1400 mg/litre, potassium nitrate (KNO ₃) 300-500 mg/litre, calcium chloride (CaCl2H ₂O) 150-300 mg/litre, magnesium sulfate (MgSO ₄7H ₂O) 40-100 mg/litre, potassium dihydrogen phosphate (KH ₂PO ₄) 200-400 mg/litre, ammonium dihydrogen phosphate (ADP) (NH ₄H ₂PO ₄) 50-100 mg/litre, sulfuric acid argon iron (FeSO ₄7H ₂O) 13.9 mg/litre, disodium ethylene diamine tetraacetate (Na ₂EDTA) 18.6 mg/litre, potassium iodide (KI) 0.83 mg/litre, boric acid (H ₃BO ₃) 3.1 mg/litre, manganese sulphate (MnSO ₄H ₂O) 8 mg/litre, zinc sulphate (ZnSO ₄7H ₂O) 8.3 mg/litre, sodium molybdate (Na ₂MoO ₄2H ₂O) 0.25 mg/litre, copper sulphate (CuSO ₄5H ₂O) 0.25 mg/litre, cobalt chloride (COCl ₂6H ₂O) 0.025 mg/litre, inositol 50 mg/litre, nicotinic acid 0.5 mg/litre, glycine 1 mg/litre, thiamine hydrochloride 0.5 mg/litre, hydrochloric acid adjoin plain 0.5 mg/litre of trembling, sucrose 20 mg/litre, agar 5.5 mg/litre, PH5.8, and explant (axillalry bud) is cultivated and differentiated the bud of growing thickly in 25 days on the medium of additional plant hormone BA2 mg/litre, ZE1 mg/litre, IAA0.2 mg/litre.Successive transfer culture, the sprout tuber that will grow thickly are transferred on the medium of additional plant hormone BA0-25 mg/litre, IAA0.5 mg/litre, and the interior culture of bottle grows the sturdy no offspring of 3-5 strain after 20 days.Again to no offspring root induction, no offspring is transferred on the medium of additional plant hormone NAA0.05 mg/litre BA0.025 mg/litre, the interior little strain of bottle grows the sturdy root of 2-5 bar from basal part of stem after 20-25 days, the root length of plantlet is taken out from medium during to 1-1.5 centimetre, after running water (or clear water) cleans, direct transplanting in the compost, is put the greenhouse and is supported seedling in the basin, moves on to outdoor management when plant is long again after to 15-20 centimetre.[transplant the back and grow new cauline leaf after 15-20 days season in spring and autumn, adopts winter the heavy caliber flowerpot to contain the intensive transplanting plantlet of compost, puts culturing room or greenhouse heating (15-25 ℃) and support seedling, and survival rate is more than 80%].The living plant of 2-3 just manages to make do flower bud to bloom under suitable condition of pot.

Claims

1, the stripped rapid breeding method of a kind of plum blossom is characterized in that adopting the autogamy medium that hangs down inorganic salt concentration, and plant hormone is combined as compound, and explant adopts the elite stand axillalry bud of growing up, vegetative progeny good reliability.

2, according to claim 1, it is characterized in that the medium of autogamy is made up of following composition: ammonium nitrate (NH ₄NO ₃) 1200～1400 mg/litre, potassium nitrate (KNO ₃) 300～500 mg/litre, calcium chloride (CaCl2H ₂O) 150～300 mg/litre, magnesium sulfate (MgSo ₄7H ₂O) 40～100 mg/litre, potassium dihydrogen phosphate (KH ₂Po ₄) 200～400 mg/litre, ammonium dihydrogen phosphate (ADP) (NH ₄H ₂Po ₄) 50～100 mg/litre, sulfuric acid argon iron (FeSo ₄7H ₂O) 13.9 mg/litre, disodium ethylene diamine tetraacetate (Na ₂EDTA) 10.6 mg/litre, potassium iodide (KI) 0.83 mg/litre, boric acid (H ₃Bo ₃) 3.1 mg/litre, manganese sulphate (MnSO ₄H ₂O) 8 mg/litre, zinc sulphate (ZnSo ₄7H ₂O) 8.3 mg/litre, sodium molybdate (Na ₂MoO ₄2H ₂O) 0.25 mg/litre, copper sulphate (CuSO ₄5H ₂O) 0.25 mg/litre, cobalt chloride (CoCl ₂6H ₂O) 0.025 mg/litre, inositol 50 mg/litre, nicotinic acid 0.5 mg/litre, glycine 1 mg/litre, thiamine hydrochloride 0.5 mg/litre, pyridoxine hydrochloride 0.5 mg/litre, sucrose 20 mg/litre, agar 5.5 mg/litre, PH5.8.

3, according to claim 1, cultivation is carried out on the medium of additional BA2 mg/litre, ZF1 mg/litre, IAA0.2 mg/litre at the beginning of it is characterized in that explant (axillalry bud), the successive transfer culture additional BA0.25 mg/litre of bud of growing thickly is induced seedling take root additional BA0.05 milligram, BA0.025 mg/litre.