| Sophora flavescens Ait.is a perennial deciduous shrub or herbaceous plant of the genus Leguminosae.It is a commonly used Chinese medicinal material.The wild resources of Sophora flavescens are widely distributed.However,in recent years,the market supply is mainly artificial cultivation.The seeds of Sophora flavescens have hardseed,slow swelling rate,low germination rate,slow germination and untidy emergence.At present,there are few studies on hardseed,and there are few studies on the hard and hardseed genes of Sophora flavescens.Therefore,it is of great significance to study the characteristics and mechanisms of the hardseed nature of Sophora flavescens.The seed morphological characteristics of the seeds of Sophora flavescens and the seed ginseng of different stages were observed.The hard and germination characteristics of Sophora flavescens seeds were studied by concentrated sulfuric acid,friction and hot water.The paraffin section method was used to study the tissue anatomy of Sophora flavescens seed coats in different periods,and the relationship between hardseed and seed coat structure was discussed.Based on the Gm Hs1-1 gene of soybean hardseed gene,the Sf Hs1-1 gene was controlled by PCR cloning,and the expression level of Sf Hs1-1 was detected by q RT-PCR.Then the bioinformatics tool was used to test the hardness.Genes were analyzed to explore the role of hardseed genes in dormancy.The results in the study are showing as follows:1.The seed of Sophora flavescens is elliptical or obovate,the seed coat is light brown or tan,the surface is smooth and shiny,and the umbilical cord is obvious,and the texture is hard.The hardseed rate is over 90%,the seed size varies from seed source to seed,the seed length is 4.50-5.88 mm,the width of the seed is 3.46-4.43 mm,and the aspect ratio is 1.30-1.43.The seed has a thousand-grain weight of 41.18-58.90 g.The viability of the seed of the different provenances varies greatly,with a minimum of 38% and a maximum of 98%.2.Sophora flavescens has a high hardseed rate,the hardseed rate is at least 87%,up to 99%;different methods of breaking can significantly reduce the hardseed rate.3.Taking germination rate as a reference,concentrated sulphuric acid treatment,friction treatment and hot water soaking can significantly improve seed germination rate compared with untreated,and the optimal treatment of different provenances is different.4.Sophora flavescens seed coat is composed of stratum corneum,palisade layer,bone stone cell,thick-walled cell layer and parenchyma cell layer.It has typical leguminous seed coat structure characteristics and is a relatively complex seed coat structure;The development of the seed coat of Kushen is thickened,the stratum corneum is thickened,and the cells of the palisade are arranged more closely and the proportion is significantly increased.5.The cloned hard stalk gene SfHs1-1 has an open reading frame of 1386 bp and encodes 461 amino acids.The amino acid sequence contains one conserved domain and is a family of alkaline phosphatases.Through software analysis,Sf Hs1-1 has a molecular formula of C2360H3671N647O662S15,a molecular weight of 52267.15 k D,an isoelectric point of 9.21 and a stability coefficient of 33.94.It is a stable hydrophilic protein;it forms a random coil-β-fold in space.Complex configuration such as random coil-α-helix-free coil,mainly located in the plasma membrane;the expression level is different in the seed coat in different periods,and the whole body first rises and then decreases,and the expression in the fifth week The highest volume provides the basis for further research on its functions in the future. |
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