In-vitro identification method for gramineal self-incompatibility phenotype

An identification method and affinity technology, applied in the field of plant biology, can solve problems such as time-consuming, complex genetic background, and large workload, and achieve the effects of simple and easy-to-master methods, controllable experimental conditions, and improved efficiency.

Inactive Publication Date: 2018-11-30
JIANGSU UNIV
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0003] But on the other hand, self-incompatibility plants have been sexually reproduced by hybridization in nature for a long time, and the heterozygosity of individual genomes is high, and the genetic background is complex. Since it is impossible to obtain self-bred pure lines, traits segregate in the hybrid F1 generation , not only reduces the performance of its heterosis, it is not suitable for large-scale production of hybrid seeds to promote planting, but also greatly limits the development of genetic research such as gene mapping, gene map cloning and whole genome sequencing, hindering further genetic improvement, Therefore, it is necessary to find a way to overcome its self-incompatibility
[0004] In the hybrid breeding and genetic research of Miscanthus and other plants, it is often necessary to mea

Method used

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  • In-vitro identification method for gramineal self-incompatibility phenotype

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Experimental program
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Effect test

Embodiment 1

[0049] Source: 20 M. sinensis genotypes preserved in Jiangsu University, numbered Ms01~Ms20.

[0050] 1. Inflorescence arrangement: select the spikes that will open the next day from plants of different awn genotypes, cut off the inflorescences with branch shears, retain the stalks about 1 meter in length at the bottom, and place a small amount of the inflorescences that are already open. The spikelets are trimmed away.

[0051] 2. Artificial flower promotion: insert the arranged inflorescence branches into the Hoagland nutrient solution, and place them in complete darkness at room temperature for 8-12 hours for flower promotion.

[0052] 3. In vitro culture of pistil: Take the mature spikelet that will open the next day from the inflorescence of the same awn genotype plant, peel off the inner and outer lamina of the spikelet with pointed tweezers under an OLYMPUS stereo microscope to expose the pistil, and then use it A sharp surgical blade cuts the intact pistil from the base of t...

Embodiment 2

[0058] Source: M. sinensis Ms344 and different individual plants of the progeny population of cross-crossing with Ms425.

[0059] 1. Inflorescence arrangement: select the spikes that will open the next day from the progeny plants of the male parent, and cut the inflorescences with branch shears. Keep a stalk about 1 meter in length at the bottom, and place a small amount on the inflorescence The open spikelets are trimmed away.

[0060] 2. Artificial flower urging: insert the above-mentioned inflorescence branches into the Hoagland nutrient solution, and place them in complete darkness at room temperature for 8-12 hours for flower urging treatment.

[0061] 3. In vitro culture of pistil: Take the mature spikelet that will open the next day from the inflorescence of the Ms344 plant, peel off the inner and outer lamina of the spikelet with pointed tweezers under the OLYMPUS stereo microscope to expose the pistil, and then use a sharp operation Cut the intact pistil from the base of th...

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Abstract

The invention belongs to the technical field of plant biology, and discloses an in-vitro identification method for gramineal self-incompatibility phenotype, which comprises the following steps: (1) inflorescence arrangement: the inflorescences of a paternal plant are pruned at the full-bloom stage, removing bloomed spikelets; (2) artificial flower forcing: the inflorescences are inserted into Hoagland nutrient solution, and flower forcing treatment is carried out under a completely dark condition for 8 to 12 hours; (3) in-vitro pistil culture: mature spikelets are chosen from the maternal inflorescences, and the pistils are intactly cut off under a stereoscopic microscope and inserted into 0.5 to 1 percent of agarose solid medium for in-vitro culture; (4) indoor pollination: in the morningof the next day, paternal pollen is used for pollinating the pistils; (5) pistil softening: the pollinated pistils are softened with 8 to 10mol/L of NaOH for 3 to 4 hours; (6) pistil dyeing: the softened pistils are dyed with 0.1 to 0.5 percent of aniline blue; (7) microscopic observation: the growth of the pollen tubes on the pistil stigmas are observed in an inverted fluorescence microscope. The method is simple and easy to master, the accuracy is high, and the method is suitable for being applied in the genetic analysis and crossbreeding research of miscanthus.

Description

Technical field [0001] The invention belongs to the field of plant biotechnology, and specifically relates to an in vitro identification method for the self-incompatibility phenotype of the Gramineae. Background technique [0002] Miscanthus (Miscanthus) is an important class of C4 gramineous energy grass and forage grass. It has the characteristics of high biomass production and strong stress resistance. It is naturally distributed in East Asia and Southeast Asia. my country mainly has awn (M.sinensis), M. floridulus (M. floridulus), Di (M. sacchariflorus), South Di (M. lutarioriparius) and other species, they all have strict self-incompatibility (Self-incompatibility, SI), that is, self-incompatibility. Self-incompatibility is of great significance for plants to avoid inbreeding and maintain genetic diversity. It is also one of the main ways to use plant heterosis. Self-incompatibility is used as the female parent to make it compatible with other self-incompatibility. Crossing ...

Claims

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Application Information

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IPC IPC(8): A01H1/02A01H1/04
CPCA01H1/02A01H1/04
Inventor 蒋建雄孙建中高璐王永丽武艳芳李霞
Owner JIANGSU UNIV
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