Method for improving germination rate of cultivation tobacco-Nicotiana stocktonii crossbred pollen

A technology of pollen germination and Tuoketong’s technology, which is applied in the field of agricultural biology, can solve the problems of large differences in pollen germination rate, abnormal microspore development, and low germination rate, and achieve high pollen germination rate, simple operation, and low cost. Effect

Inactive Publication Date: 2015-12-09
YUNNAN UNIV
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Problems solved by technology

For example, the cross-pollination experiment of ♀Nta(gla.)SK326×♂N.stocktonii was carried out in the Yuxi experimental field in Yunnan in winter. Due to the abnormal development of microspores due to low temperature, the pollen germination rate on the stigma after cross-pollination was lower than 15%. ♀Nta(gla.)SK326×♂N.stocktonii cross-pollination experiments were carried out in the base, and the microspore development process of the male parent was normal, but the pollen germination rate on the stigma was very different after pollination in different time periods

Method used

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] The concrete steps of this method are as follows:

[0015] 1. Seedlings were raised in September 2012 and transplanted in October, namely the female parent Nta(gla.) SK326 and the male parent N. stocktonii. The flowering period was January-February 2013. The mature pollen of the male parent, N. stocktonii, was collected during the flowering period. In 10:00-12:00, 12:00-14:00, 14:00-16:00, 16:00-18:004 time periods, the mature pollen of the male parent N.stocktonii was used The flowers of the cultivated tobacco, namely the female parent Nta(gla.) SK326, were pollinated at the beginning stage, and were isolated by bagging after pollination.

[0016] 2. Take off the nylon mesh bag 12 hours after pollination, pick off the flowers and peel off the pistil, store it in FAA fixative solution, place it at 4°C for more than 2 hours, discard the fixative solution; soften with NaOH for 120 minutes, and dye with 0.1% water-soluble aniline blue After more than 2 hours, take pictur...

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Abstract

The invention discloses a method for improving a germination rate of cultivation tobacco-Nicotiana stocktonii crossbred pollen and belongs to the technical field of agricultural living things. The method comprises a, collecting mature pollen of Nicotiana stocktonii as a male parent in a flowering phase, and in four time periods of 10:00-12:00, 12:00-14:00, 14:00-16:00 and 16:00-18:00, respectively carrying out pollination on female parent stigmata in an early flowering stage with the collected mature pollen, b, coating the female parent stigmata by CaCl2 solutions with concentrations of 10mg / L, 30mg / L and 60mg / L, and in a time period of 10:00 AM-12:00 AM, carrying out pollination on the CaCl2-coated female parent stigmata in an early flowering stage with Nicotiana stocktonii mature pollen, c, coating the female parent stigmata in an early flowering stage with aminoethoxyvinyl glycine solutions with volume fractions of 0.0001%, 0.001% and 0.1% as ethylene removal agents, and in a time period of 10:00 AM-12:00 AM, carrying out pollination on the aminoethoxyvinyl glycine-coated female parent stigmata in an early flowering stage with male parent mature pollen, and d, after pollination for 12h, carrying out sampling, carrying out tabletting, taking pictures by a fluorescent microscope and making statistics of a pollen germination rate, wherein the above female parent is cultivation tobacco. The method can be operated simply, has a low cost and a high pollen germination rate and provides effective guidance for improving stigma cross compatibility.

Description

Technical field: [0001] The invention relates to a method for improving the germination rate of hybrid pollen of cultivated tobacco and Nicotiana stocktonii, belonging to the field of agricultural biotechnology. Background technique: [0002] Distant hybridization between plant species can break the boundaries between species to a certain extent, and recombine the beneficial traits accumulated by two species through long-term evolution to form new yield, quality traits and resistance to diseases, insects, cold, drought, Waterlogging and other stress resistance (tolerance), etc. Distant hybridization has played an important role in the production and breeding of plants such as wheat, cotton, rice and tomato. However, in many crop cross-breeding combinations, interspecific genetic background differences and reproductive isolation often lead to hybrid incompatibility. This phenomenon is also commonly found in Nicotiana plants, including hybrid incompatibility before fertiliza...

Claims

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Application Information

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IPC IPC(8): A01H1/02A01G7/06
Inventor 陈穗云廖菊够李永平马文广
Owner YUNNAN UNIV
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