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In-vitro regeneration culture method of tree peonies

An in vitro regeneration and peony technology, which is applied in plant regeneration, horticultural methods, botanical equipment and methods, etc., can solve the problems of general effect and slow in vitro regeneration of peony, and achieve the effect of promoting growth.

Active Publication Date: 2017-05-10
南京康之源农业科技发展有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve the above-mentioned problems of slow regeneration of tree peony in vitro and general effect, the present invention forms hypocotyls through the regeneration of mature embryos, and makes the hypocotyls suddenly relieved under the stimulation of plant growth regulator 6-BA and IBA and the osmotic stress of PEG6000 Under these conditions, adventitious buds were produced directly without callus, and the improved MS medium was additionally added with proline and riboflavin during rooting to achieve rapid propagation of peony, and the rooting rate reached 70%. , transgenic and other genetic manipulations to induce new varieties lay the foundation

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Step 1, after sterilizing the peony seeds, take the mature embryos and inoculate them on 1 / 2MS medium for cultivation;

[0033] Step 2. After the mature embryo germinates, take the young hypocotyl, cut it into 0.4cm long, inoculate it in PEG6000 containing 4% (W / V) and add 1.5 mg / L 6-BA, 0.2mg / L Cultured on the modified MS medium of LIBA and 3% (W / V) sucrose for 15 days until the hypocotyl incision was enlarged;

[0034] Step 3: Inoculate the enlarged hypocotyls at the incision on the modified MS medium supplemented with 1.5 mg / L 6-BA and 0.2 mg / L IBA for 14 days until the hypocotyls are directly differentiated at the incision Adventitious buds of peony;

[0035] Step 4. When the adventitious buds of peony grow to 3cm, cut off the adventitious buds of peony from the root, and inoculate them with 0.2 mg / L NAA, 0.2 mg / L IBA, 50 mg / L proline and 0.1 mg / L nuclear yellow rooting on the improved MS medium of Su, that is, complete the in vitro regeneration of peony.

[0036...

Embodiment 2

[0040] Step 1, after sterilizing the peony seeds, take the mature embryos and inoculate them on 1 / 2MS medium for cultivation;

[0041] Step 2. After the mature embryo germinates, take the young hypocotyl, cut it into 0.6cm long, inoculate it in PEG6000 containing 4% (W / V) and add 2.5 mg / L 6-BA, 0.5 mg / L Cultured on the modified MS medium of LIBA and 3% (W / V) sucrose for 25 days until the hypocotyl incision was enlarged;

[0042] Step 3: Inoculate the enlarged hypocotyls at the incision on the modified MS medium supplemented with 2.5 mg / L 6-BA and 0.5 mg / L IBA for 30 days until the enlarged hypocotyls are directly differentiated into Adventitious buds of peony;

[0043] Step 4. When the adventitious buds of peony grow to 4cm, cut off the adventitious buds from the root and inoculate them in the seedlings supplemented with 0.2 mg / L NAA, 0.2 mg / L IBA, 100 mg / L proline and 3 mg / L riboflavin Rooting on 1 / 2 of the modified MS medium, that is to complete the in vitro regeneration o...

Embodiment 3

[0048] Step 1, after sterilizing the peony seeds, take the mature embryos and inoculate them on 1 / 2MS medium for cultivation;

[0049] Step 2. After the mature embryo germinates, take the young hypocotyl, cut it into 0.5cm long, inoculate it in PEG6000 containing 4% (W / V) and add 2 mg / L 6-BA, 0.35mg / L Cultured on the modified MS medium of LIBA and 3% (W / V) sucrose for 20 days until the hypocotyl incision was enlarged;

[0050] Step 3: Inoculate the enlarged hypocotyls at the incision on the modified MS medium supplemented with 2 mg / L 6-BA and 0.35 mg / L IBA for 22 days until the enlarged hypocotyls are directly differentiated from the incision. Adventitious buds of peony;

[0051] Step 4. When the adventitious buds of peony grow to 3.5cm, cut off the adventitious buds from the root and inoculate them in the seedlings supplemented with 0.2 mg / L NAA, 0.2 mg / L IBA, 75 mg / L proline and 1.4 mg / L nucleus. Rooting on the modified MS medium of flavin, that is to complete the in vitro...

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Abstract

The invention provides an in-vitro regeneration culture method of tree peonies. The in-vitro regeneration culture method comprises the following steps: after sterilizing tree peony seeds, taking mature embryos and inoculating the mature embryos on a 1 / 2MS culture medium for culturing; after the mature embryos sprout, taking tender hypocotyls; inoculating the tender hypocotyls into an improved MS culture medium and culturing for 15 to 25 days until notches of the hypocotyls are expanded; inoculating hypocotyl subcultures with the expanded notches into the improved MS culture medium for 14 to 30 days until tree peony adventitious buds are directly differentiated on the expanded notches of the hypocotyls; after the tree peony adventitious buds grow to 3cm to 4cm, cutting off the tree peony adventitious buds from roots and inoculating the tree peony adventitious buds on the improved MS culture medium for rooting, so as to finish in-vitro regeneration of the tree peonies. According to the in-vitro regeneration culture method of the tree peonies, provided by the invention, the adventitious buds can be directly produced by the hypocotyls without calluses under the condition of suddenly reliving PEG6000 (Polyethylene Glycol 6000) osmotic stress and increasing the concentrations of Ca<2+> and microelements Mn<2+>, Zn<2+>, Mo<6+> and Cu<2+>, and a callus process is omitted, so that the problems that the adventitious buds are relatively difficult to differentiate by the calluses and the adventitious bud differentiation rate is commonly relatively low are solved.

Description

technical field [0001] The invention relates to the technical field of peony tissue culture, in particular to a tissue culture method for peony regeneration in vitro. Background technique [0002] Peony, a plant of the genus Paeoniae in the family Paeoniaceae, is a traditional famous flower in my country. It is used for both ornamental and medicinal purposes. In recent years, peony varieties such as 'Fengdanbai' have been found to have high oil content and high quality. They are positioned as high-end edible oils. It is widely planted in Luoyang, Henan, and even the whole country, such as Tibet. The large-scale planting of peonies not only drives the tourism industry of one side, but also drives the rapid development of the local economy. Therefore, it is particularly important to study its quality breeding, improve its quality, and develop our advantageous industries. [0003] However, due to reasons such as long childhood and complex genetic background of peony, tradition...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 张改娜史国安侯典云
Owner 南京康之源农业科技发展有限公司
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