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| 1 | Molecular Marker Techniques Using Single Primers and Their Advances显示文摘Molecular marker techniques have been widely applied in the fields of genetic diversity analysis,germplasm resources identification,molecular fingerprint and genetic linkage map construction,QTL mapping and molecular assisted breeding.On the basis of stating the concept of molecular marker techniques based on single primer amplification reactions,this study focused on the sorting and induction of single-primer molecular marker techniques,and expounded their derivative development.Finally,the application prospect and future expectation of single-primer molecular marker techniques were described in detail.The purpose of this study was to clarify the types of molecular marker techniques based on single primer amplification reactions,so that researchers can quickly and conveniently select molecular marker techniques according to their own specific scientific research conditions. | Junxian LIU Jing LIU Jing JIANG Zhuqiang HAN Xiumei TANG Lihang QIU Ruichun ZHONG Liangqiong HE Haining WU Zhipeng HUANG Ronghua TANG Faqian XIONG | 2021 | Agricultural Biotechnology2021，10，2： | 0 |
| 2 | A Non-toxic and Efficient Method for Extracting DNA and RNA from Peanut显示文摘[Objectives]To establish a non-toxic and efficient method for extracting DNA and total RNA from peanuts and laying a solid foundation for the molecular biology study of peanuts.[Methods]Based on the principle and method of purifying nucleic acids by silica gel adsorption at high salt and low pH condition,a non-toxic and efficient method to extract peanut DNA and total RNA using cetyltrimethyl ammonium bromide(CTAB)extraction solution was designed.The quality and purity of nucleic acids were detected by agarose gel electrophoresis and nucleic acids protein analyzer,respectively.The quality of DNA was further verified by enzyme digestion and PCR amplification using molecular marker techniques.The quality of total RNA was further verified by reverse transcription(RT)-PCR of actin gene and cDNA-SCoT gene differential display technique.[Results]The agarose gel electrophoresis test showed that the peanut DNA extracted by a low-toxic and effective method is free of contamination and degradation.Through the detection by the nucleic acid protein analyzer,the DNA concentration,yield,A260/A280 and A260/A230 of 5 peanut varieties were 419.6-498.2 ng/μL,20.98-24.91μg/g,1.89-1.96 and 2.03-2.28,respectively.The DNA was of high quality and can be completely digested by EcoRI restriction enzymes,and also can be used for SCoT and SRAP molecular marker technology analysis.The RNA extracted from different tissues of peanuts showed no visible DNA bands by non-denaturing agarose gel electrophoresis.The separated 28S bands were brighter than 18S.The ratio of A260/A280 and A260/A230 showed that the RNA quality was good and can be used for reverse transcription,RT-PCR of actin gene and amplification of cDNA-SCoT gene differential display technique.[Conclusions]This experiment established a low-toxic and effective method for extracting DNA and total RNA from peanuts.Compared with traditional methods,this method is more time-saving and cheaper than commercial kits.The most important point is that this method does not use toxic reagents such as phenol,chloroform and isopropanol.Thus,it is expected to be widely applied in molecular biology research. | Jing LIU Congcong WANG Jing JIANG Zhuqiang HAN Xiumei TANG Lihang QIU Liangqiong HE Ruichun ZHONG Zhipeng HUANG Haining WU Ronghua TANG Junxian LIU Faqian XIONG | 2021 | Asian Agricultural Research2021，13，1： | 0 |
| 3 | Effect of Gibberellin,Light and Genotype on Somatic Embryogenesis and Plantlet Regeneration of Peanut显示文摘[Objective]Peanut in vitro regeneration system was optimized to provide technical support for its genetic transformation and mutagenesis.[Method]The effect of gibberellin,light and genotype on somatic embryogenesis and plantlet regeneration of peanut was studied using Guihua peanut cultivar as materials,leaflets as explants,MS+10 mg/L 2,4-D as somatic embryo induction medium,and MS+3 mg/L 6-BA+0.8 mg/L NAA+(0-15 mg/L)GA3as plantlet induction medium.[Result]The somatic embryo induction rate of five peanut varieties under light condition of light:dark=14 h:10 h was significantly higher compared with that under dark condition by 7.5-37.5 percent points.Among different peanut varieties,Guihua 26 represented the highest somatic embryo induction rate of62.8%,while Guihua 833 represented the lowest somatic embryo induction rate of 21.7%.Adding 5-15 mg/LGA3in plantlet induction medium was conducive to improving the induction rate of plantlets derived from somatic embryos.With addition of 5 mg/L GA3,Guihua 26 and Guihua 771 represented the highest plantlet induction rate of 42.8%and 35.3%,respectively;with addition of 15 mg/L GA3,Guihua 836,Guihua 1026 and Guihua 833 represented the highest plantlet induction rate of 38.7%,33.3%and 26.4%,respectively.Among different combinations of peanut variety and GA3concentration,plantlet induction rate reached the highest in the combination Guihua 26+5 mg/L GA3and reached the lowest in the combination Guihua 836+15 mg/L GA3.[Conclusion]Appropriate light conditions are conducive to peanut somatic embryogenesis in vitro.Adding GA3in plantlet induction medium is conducive to promoting plantlet regeneration.Guihua26 and Guihua 836 are the best peanut varieties among the experimental genotypes for somatic embryogenesis and plant regeneration. | Jing JIANG Faqian XIONG Xiumei TANG Ruichun ZHONG Liangqiong HE Zhong LI Zhuqiang HAN Ronghua TANG | 2013 | Agricultural Biotechnology2013，2，6： | 0 |
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