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Improvement of isolated microspore culture of barley (Hordeum vulgare L.): the effect of floret co-culture

文献类型： 外文期刊

作者： Lu, Ruiju 1 ; Wang, Yifei 1 ; Sun, Yuefang 1 ; Shan, Lili 1 ; Chen, Peidu 3 ; Huang, Jianhua 1 ;

作者机构： 1.Shanghai Acad Agr Sci, Biotech Res Inst, Shanghai 201106, Peoples R China

2.Shanghai Key Lab Agr Genet & Breeding, Shanghai 201106, Peoples R China

3.Nanjing Agr Univ, Natl Key Lab Crop Genet & Germplasm Enhancement, Jiangsu 210095, Peoples R China

关键词： multicellular structures (MCS);embryo-like structures (ELS);plant regeneration;androgenesis;TRITICUM-AESTIVUM L;WHEAT;EMBRYOGENESIS;REGENERATION

期刊名称：PLANT CELL TISSUE AND ORGAN CULTURE （ 影响因子：2.711； 五年影响因子：2.73 ）

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收录情况： SCI

摘要： Barley microspores from five field-grown breeding lines were isolated using an ultra-speed blender and the effect of co-culture with young florets was investigated. Floret co-culture in the induction stage increased the formation of MCS, ELS and green plant regeneration. The florets of teraploid plant were more effective than ones of diploid plant. For line S23, co-culture with florets from tetraploid plants gave rise to 2.6 and 7.8 times more MCS and ELS, respectively, than non-co-culture control, whereas co-culture with florets from diploid plants resulted in 1.8 and 6.1 times more MCS and ELS, respectively, than non-co-culture control (Table 2). Florets subjected to cold treatment for 10-20 days induced a greater response than fresh ones, and florets with uninucleate microspores surpassed binucleate microspores. For microspores culture from 15-day cold pre-treated spikes, 93A floret co-culture gave rise to 3.6 and 6.8 times more MCS and ELS, respectively, than the non-co-cultured control, while SD1 floret co-culture resulted in 1.9 and 4.0 times more, respectively. Similarly, for microspore culture from 20-day cold pre-treated spikes, 93A floret co-culture gave rise to 2.6 and 5.1 times more MCS and ELS, respectively, than non-co-cultured control, while SD1 floret co-culture resulted in 1.5 and 3.0 times more, respectively (Table 3). Some microspores formed dense MCS that did not develop further. Compared with the control, floret co-culture resulted in less dense MCS formation, indicating that the isolated florets were beneficial to the normal development of MCS. Floret co-culture was only effective when the spikes were cold pre-treated before microspore isolation. Spike cold pre-treatment before microspore preparation was crucial for dedifferentiation of cultured isolated microspores, and this could not be replaced by floret co-culture. It is postulated that the florets provided essential substances for in vitro cultured isolated microspores to undergo dedifferentiation and embryogenesis. Both the genotype selection and the physiological status (developmental status and cold treatment) adjustment of the florets for co-culture could improve barley microspore culture. Compared with ovary co-culture, floret co-culture is more efficient. The technique is of simple application in breeding programs and can be a solution for coping with recalcitrant genotypes and or plant donor condition.

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