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To obtain the superior genotype of HvHinb-1 gene related to the hardness of hulless barley grains, two hulless barley varieties with significant differences in hardness were seleeted: Suma (hardness value: 14.66±1.55) and Nanmulin (hardness value: 21.42±1.83). The HvHinb-1 gene sequence through PCR were doned to analyzed the sequence characteristics, protein physicochemical properties, and expression patterns of different genotypes, and explored their roles in the formation of hulless barley grain hardness. The distribution and genotype of HvHinb-1 in 21 hulless barley varieties with different hardness were analyzed, and the expression of HvHinb-1 in different stages of grain development (early, middle and late milk ripening) was detected by real-time quantitative PCR (qRT-PCR).The results showed that the HvHinb-1 was cloned from both the Suma and Nanmulin, with a sequence identity of 98.64%, and two amino acid differences. HvHinb-1 is a hydrophilic basic protein with an isoelectric point of 8.69 and a molecular weight of 16.12 to 16.14 kDa, primarily composed of α-helices and random coils. Phylogenetic analysis indicated that hulless barley HvHinb-1 has the closest genetic relationship to wheat. In the preliminary identification of 12 samples of low-hardness hulless barley (hardness value range: 9.04-15.04), the amino acid sequence at the mutation sites 78 and 94 was H, and in 9 samples of high-hardness hulless barley (hardness value range: 18.93-22.76), the amino acid sequence at the mutation sites 78 and 94 was Q. qRT-PCR analysis showed that in early milk ripening grains, the expression level of HvHinb-1 was significantly higher in Suma than that in Nanmulin, and in mid-milk ripening, it was extremely higher in Nanmulin than in Suma, suggesting that this period may be crucial for grain hardness formation.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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