| In order to find out the difference of egg quality between Green-shell eggs and white-shell eggs among different genotypes of Chishui Black-bone Chicken,And further improve and perfect the research content of egg quality of Chishui Black Bone Chicken.At the same time,in order to find out the reason why Chishui Black-bone Chicken exists in different genotypes affecting the green depth of Green Eggshell eggshell,and verify the reasons.Chishui Black Bone Chicken was taken as the research object in this experiment,and egg quality analysis,transcriptome sequencing analysis,expression analysis of liver and eggshell gland between different eggshell green depths of different genotypes in different periods were used as three aspects to study Chishui Black Bone Chicken.It is hoped that the experiment can provide data basis for further research on the green depth of Chishui Black-Boned Chicken’s eggshell,and provide effective theoretical support for the conservation and utilization of Chishui BlackBoned Chicken,so as to improve the production efficiency of Chishui Black-Boned Chicken.The main contents of the experiment are as follows:1: 8 months of Chishui silky fowl hens 314 were chosen as research subjects.First,according to the genotyping results of SLCO1B3,it was divided into three types: green eggshell homozygous(SL/SL),green eggshell heterozygous(SL/W)and white eggshell homozygous(W/W).Then,60 eggs of each genotype were collected to determine the egg weight,shell color,egg shape index,shell strength,protein height,yolk color,Huff value,yolk weight,the content of biliverdin and protoporphyrin in eggshell.(Result)Finally,the results are as following: There was no significant difference in eggshell index and the egg yolk color between the 3 genotypes in the egg quality study(P>0.05),and all of them were between 1.3-1.35,the eggshell strength of green eggshell homozygous type was significantly lower than that of green eggshell heterozygous type and white eggshell homozygous type(P<0.05),the egg weight of green eggshell heterozygous type was significantly higher than that of white eggshell homozygous type and green eggshell homozygous type(P<0.05),the protein heights of green eggshell homozygous type and white eggshell homozygous type were significantly higher than those of green eggshell heterozygous type(P<0.05),the egg yolk weight of green eggshell homozygous type was significantly lower than that of green eggshell heterozygous type(P<0.05),and the egg yolk weight of white eggshell homozygous type was between the two genotypes and was no significant difference from the other two genotypes(P>0.05),,the Huff values of green eggshell homozygous type and white eggshell homozygous type were significantly higher than those of green eggshell heterozygous type(P<0.05).In the study of the content of biliverdin and protoporphyrin in eggshell,the content of biliverdin in eggshells of green eggshell homozygous type eggshells was found to be significantly higher than that of green eggshell heterozygous type eggshells and white eggshell homozygous type eggshells(P < 0.05),and the green eggshell heterozygous type eggshells was significantly higher than that in white eggshell homozygous type eggshells.The content of protoporphyrin in white eggshell homozygous type eggshells was significantly higher than that in eggshell heterozygous type eggshells and green eggshell homozygous type eggshells(P<0.05),and the green eggshell heterozygous type eggshells was significantly higher than that in green eggshell homozygous type eggshells(P<0.05).(Conclusion)The results showed that the egg quality of green eggshell homozygous type Chishui silky fowl was better than that of green eggshell heterozygous type,the market value was higher than that of white eggshell homozygous type,and the green eggshell homozygous type Chishui silky fowl had higher development and utilization value.2: 360 days of Chishui silky fowl hens 300 were chosen as research subjects,and according to the green depth of eggshell and genotyping results of SLCO1B3,they were divided into six types: homozygous dark green,homozygous deep green,homozygous light green,heterozygous dark green,heterozygous deep green and heterozygous light green.There are 3 in each types and 18 in all were selected as the research objects,Chishui silky fowl were slaughtered 2.5 H-3 h before laying eggs,and their shell glands were taken for transcriptome sequencing.The results were as follows: 843,052,836 Reads(sequencing sequence)were obtained,the contaminated Reads were removed,the low-quality Reads were removed,and the Reads containing more than 5% N were removed.Finally,801,602,652 Clean Reads(high-quality sequence)were obtained.More than 40,000,000 Reads were detected in a single sample during the sequencing process,and the proportion was over 93.00%.The transcriptome sequencing analysis showed that there were three differentially expressed genes in different eggshell green depths of different genotypes.The up-regulated gene was CNTN5 gene and SPIA1 gene,and the down-regulated gene was CHRDL1 gene.The screening of differentially expressed genes was helpful to control and improve the eggshell quality of Chishui silky fowl in the later stage and promote the development of Chishui silky fowl.3: 180 days,360 days and 540 days of Chishui silky fowl hens who had been genotyped for green shells and classified for green depth of eggshells were chosen as research subjects,The eggshell glands and liver were taken from the eggshell 2.5 h to 3 h before laying eggs,Real-time fluorescence quantitative PCR was used to validate differentially expressed genes.The result is as follows::The expression levels of SPIA5 gene in liver and eggshell glands of Chishui silky fowl with the same genotype and different eggshell colors were dark green > light green > light green,the expression levels of CHRDL1 gene in liver and eggshell glands of Chishui silky fowl with the same genotype and different eggshell colors were dark green < light green < light green,the expression levels of CNTN5 gene in the liver and eggshell glands of Chishui silky fowl with the same genotype and different eggshell colors was close to the dark green > light green > light green,however,CNTN5 gene was irregularly expressed in the experiment,and there were errors.The expression of SPIA1 gene in eggshell gland was higher than that of CNTN5 gene and CHRDL1 gene,while that of CNTN5 gene in liver was higher than that of SPIA1 gene and CHRDL1 gene were also found.It was also found that the expression of SPIA1 and CNTN5 at 360 days was lower than that at 180 days and 540 days,while the expression of CHRDL1 at 360 days was higher than that at 180 days and 540 days.The results showed that SPIA1 gene and CHRDL 1 gene could be used as candidate genes related to the green depth of eggshell,and could be used for molecular assisted selection,so as to improve the color uniformity of eggshell and economic benefits of green shell eggs of Chishui silky fowl. |
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