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Comparative Analysis of Chromosome Sets Characteristics of Sesame Cultivated Species and Wild Species Based on FISH and GISH Technologies

MA Qin, ZHAO Ruihong, JU Ming, CHEN Chengbin, DUAN Yinghui, YANG Weifei, MIAO Hongmei, ZHANG Haiyang,

Journal of Henan Agricultural Sciences 2024, 53 (10): 48-53. DOI: 10.15933/j.cnki.1004-3268.2024.10.006

Abstract （1397）

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To reveal the evolutionary characteristics of Sesamum,explore the genome structure revolution and species evolution,and promote the development and utilization of wild resources,the cultivated species S.indicum(var.Yuzhi 11)and the 2n=26 type wild species S.alatum(var.3651)were used as experimental materials,fluorescence in situ hybridization(FISH)and genome fluorescence in situ hybridization(GISH)techniques were used to analyze the chromosome sets characteristics of sesame cultivated species and wild species.The results showed that both the cultivated species Yuzhi 11 and the wild species S.alatum(var.3651)were 2n=2x=26 karyotype;the rDNA⁃FISH hybridization results indicated that in the 13 pairs of chromosomes of the cultivated species Yuzhi 11,3 pairs of chromosomes(the 7th,8th and 9th pairs)had 45S rDNA⁃specific signals at the short arm tips,which were displayed as satellite specific chromosome.At the same time,2 pairs of chromosomes(the 5th and 11th pairs)had 5S rDNA⁃specific signals on the short arms,with the 5S rDNA and 45S rDNA signals located on different chromosomes. In the wild species S.alatum(var.3651),2 pairs of chromosomes(the 4th and 7th pairs)carried 45S rDNA hybridization signals and 1 pair of chromosome(the 4th pair)carried 5S rDNA specific signals,with the 5S rDNA and 45S rDNA signals located on the same chromosome but at different positions.This indicated that there were significant differences in chromosome characteristics between the cultivated species and the wild species S.alatum(var.3651).The GISH hybridization results showed that when using the genomic DNA of the cultivated species Yuzhi 11 and the wild species S.alatum(var.3651)as probes for self⁃hybridization,each chromosome carried hybridization signals of varying strengths,while hybridization with the other’s chromosomes showed very few hybridization signals.The cultivated species and the wild species S.alatum(var.3651)have the same number of chromosomes,but there are clear differences in the quantity,distribution of rDNA,and GISH signal sites,indicating that the 2n=26 type cultivated species and wild species have distant phylogenetic relationship.

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Screening and Analysis of SSR Markers for Evaluating Sesame Germplasm Oil Content Level

XU Fangfang, ZHENG Lei, JU Ming, MA Qin, LI Chun, DUAN Yinghui, ZHANG Xianmei, MIAO Hongmei

Journal of Henan Agricultural Sciences 2023, 52 (11): 49-56. DOI: 10.15933/j.cnki.1004-3268.2023.11.006

Abstract （709）

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In order to establish the molecular marker identification technology system related to sesame oil content level，we used 24 SSR markers closely related to sesame oil content developed by ourselves to conduct rapid PCR identification and validation analysis on 48 sesame germplasms with different oil content.The results showed that the oil content of 48 sesame germplasms ranged from 29.81% to 57.86%，with average oil content of 46.91%.The analysis of variance of SSR markers showed that 24 SSR markers showed polymorphic alleles in 48 samples，including Hs393（4/2），Hs635（2/2），Hs4082（2/1）and Hs4089（2/2）. These four SSR marker polymorphic loci were significantly（P<0.05） or very significantly（P<0.001）associated with the sesame oil content level. Among the 48 samples tested，a total of 12 samples carried excellent alleles of the four SSR markers mentioned above，with oil content ranging from 52.74% to 57.86%，all of which belonged to high oil content samples.In conclusion，these four SSR markers，Hs393，Hs635，Hs4082，and Hs4089，can be used for identifying and evaluating the oil content level of sesame germplasms and breeding materials，providing technical support for establishing and improving the molecular marker⁃assisted breeding technology system related to sesame quality.

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QTL Mapping of Seed Quality Traits Based on High‑Density Genetic Map in Sesame

CUI Chengqi, LIU Yanyang, DU Zhenwei, WU Ke, JIANG Xiaolin, ZHENG Yongzhan, MEI Hongxian

Journal of Henan Agricultural Sciences 2023, 52 (9): 66-77. DOI: 10.15933/j.cnki.1004-3268.2023.09.007

Abstract （1052）

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Sesame（Sesamum indicum L.）is an important oilseed crop for the high oil content and quality.Understanding the genetic basis of seed quality trait is essential for improving the seed quality traits of sesame. To identify the QTLs associated with the seed oil，protein，sesamin，and sesamolin content in sesame，we developed F₂，F_2:3，BC₁ and BC₁F₂ populations by crossing the Yuzhi 4 and BS lines to perform QTL mapping.A high‑density genetic map for the BC₁population was constructed using SLAF and SSR markers，and a genetic map for the F₂ population was constructed using SSR markers. In the F_2:3population，16 QTLs for the seed oil，protein，sesamin，and sesamolin content were identified and explained 5.08%—27.12% of the phenotypic variations（PVs）.Among these 16 QTLs，qOC_10‑1 was detected in two environments and explained 9.62%—27.12% of the PVs.In the BC₁ and BC₁F₂ populations，35 QTLs for the seed oil，protein，sesamin，and sesamolin content were identified.Among these 35 QTLs，the major QTLs qOC_4‑1，qOC_10‑2，and qSmin_7‑2 were detected in three environments，explaining 8.08%—12.42%，11.95%—12.60%，and 4.24%—10.56% of the PVs，respectively.Three major QTLs qSmin_8，qSmol_5‑2，and qSmol_7‑2 were identified in two environments，explaining 13.36%—26.75%，11.44%—14.33%，and 5.77%—12.38% of the PVs，respectively.By integrating the two genetic maps，we identified 10 QTL clusters.It was worth noting that loci4，loci7，loci8，and loci10 harboring at least one major QTL and associating with more than two seed quality traits were identified，and the maximum explanation rates of phenotypic variation were 12.42%，12.38%，26.75% and 27.12%，respectively.

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Screening of Nitrogen Efficient Varieties and Construction of Nitrogen Efficiency Assessment System at Seedling Stage of Sesame（Sesamum indicum L.）

ZHANG Pengyu, GAO Tongmei, SU Xiaoyu, LI Feng, WANG Dongyong, TIAN Yuan, LU Hailing, MIAO Hongmei, WEI Shuangling

Journal of Henan Agricultural Sciences 2022, 51 (6): 54-66. DOI: 10.15933/j.cnki.1004-3268.2022.06.006

Abstract （1416）

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In order to screen the sesame varieties with high nitrogen efficiency from different sesame varieties and construct an assessment system of nitrogen efficiency at seedling stage，the physiological and nitrogen efficiency traits of 18 sesame varieties at seedling stage under different nitrogen levels were measured.The variation coefficient，correlation analysis，principal component analysis，path analysis，membership function analysis and grey correlation degree analysis were used to screen nitrogen efficient varieties，determine nitrogen efficient evaluation traits，and construct nitrogen efficient evaluation system at the seedling stage of sesame.The results showed that among different varieties，there were significant differences in various traits under different nitrogen levels.Under normal and high nitrogen levels，the variation coefficients of nitrogen content and nitrogen utilization efficiency of plants showed lower than those of low nitrogen level.Under different nitrogen treatments，there was a significant or very significant positive correlation among stem dry weight，leaf dry weight，root nitrogen absorption efficiency and leaf nitrogen absorption efficiency of sesame varieties.The 18 varieties were divided into three grades by the European distance longest distance method，among them，Zhengtaizhi No.3 and Zhengzhi HL05 were nitrogen efficient varieties，Fusui Sanhe black sesame and Burmese prolific were nitrogen inefficient varieties.Under different nitrogen levels，the stem fresh weight，leaf fresh weight，leaf dry weight and stem dry weight were all highly correlated with the comprehensive score of nitrogen efficiency，which were important traits for screening of nitrogen efficient sesame varieties.Under different nitrogen levels，the two nitrogen efficient varieties showed higher advantages in root fresh weight and dry weight，and the two sesame cultivars with high nitrogen efficiency under low nitrogen level had higher taproot length，nitrogen uptake and utilization efficiency in various tissues than those under normal and high nitrogen levels.At seedling stage，low nitrogen level was more suitable for screening nitrogen efficient sesame varieties，and the nitrogen use efficiency of sesame was mainly determined by the nitrogen uptake efficiency.

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Genetic Diversity Analysis and Elite Allele Mining in Main Sesame Cultivars Released in China

LI Chun, DUAN Yinghui, JU Ming, MIAO Hongmei, DU Hua, ZHANG Haiyang

Journal of Henan Agricultural Sciences 2022, 51 (3): 55-64. DOI: 10.15933/j.cnki.1004-3268.2022.03.007

Abstract （1241）

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Researching the diversity of cultivars and mining the elite alleles underlying important traits are critical for sesame（Sesamum indicum L.）molecular breeding. In order to analyze the diversity of sesame cultivars in China，and to accelerate the progress of molecular breeding in sesame，the genome sequencing data of 705 sesame germplasms（including 95 modern sesame cultivars in China，405 landraces and 205 foreign germplasms）were analyzed in this study.As a result，12 704 SNP/InDel variants，which commonly existed in sesame lines，were identified，and by comparing the nucleotide diversity of 95 modern cultivars and 405 landraces，the cultivars were found to have a lower diversity level（0.158）than the landraces（0.246）.Further，by using the permutation test method，2 483 and 115 loci were found to possess significantly lower and higher nucleotide diversity respectively in the cultivars，and 1 290 loci were found to be significantly divergent between the cultivars and landraces using the genetic differentiation index（Fst）as the proxy. By selecting the intersection of the loci that were significantly divergent between the cultivars and landraces and the loci that possessed significantly higher nucleotide diversity in the 95 modern cultivars，80 loci were harvested. Of the 80 loci，14 were annotated to have high impact effect on gene functions，including the two loci Chr2：8176782 and Chr10：3441593，which were located in the disease resistance related genes（C_2_2.176 and C37.81）. Further analysis showed that the allele frequency of the two loci increased by 12.93 and 3.44 folds respectively in the cultivars compared with the landraces，and thus might contribute to the breeding for disease resistance. To sum up，the findings above suggest that some loci are significantly divergent between the cultivars and landraces，and the loci may relate to the improvement of sesame cultivars.

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Cloning and Prokaryotic Expression of Capsule Indehiscent Gene SiIND1 from Sesame

LIU Yanyang, WU Ke, MEI Hongxian, DU Zhenwei, CUI Chengqi, JIANG Xiaolin, ZHENG Yongzhan

Journal of Henan Agricultural Sciences 2020, 49 (5): 63-68. DOI: 10.15933/j.cnki.1004-3268.2020.05.007

Abstract （938）

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LOC105167765 belongs to KANADI (KAN) gene family,which is related with sesame capsule indehiscence.In this study,the capsule dehiscent variety Yuzhi No.11 and capsule indehiscent variety Zhengzhi InD01 were selected as experiment materials, and the SiIND1 full-length cDNAs of LOC105167765 were amplified. Bioinformatics analysis and prokaryotic expression were conducted.The results showed that the whole cDNA sequence of SiIND1 in Yuzhi No.11 was 1 320 bp and encoded 439 amino acids.The putative protein of the gene had an isoelectric point of 7.53 and a calculated molecular weight of 50.0 ku.Sequence analysis showed that the protein of SiIND1 in Yuzhi No.11 contained a SHAQKYF class MYB-like DNA binding domain similar to Arabidopsis KAN family members and shared a highly conserved GARP domain.The whole cDNA sequence of SiIND1 in Zhengzhi InD01 was 1 246 bp,which contained an ORF of 900 bp and encoded 299 amino acids.The putative protein of the gene had an isoelectric point of 8.37 and a calculated molecular weight of 32.9 ku.Compared to Yuzhi No.11,the SiIND1 gene in Zhengzhi InD01 had a deletion and generated a premature stop codon in GARP domain,which might lead to loss function of this gene.The SiIND1 gene was connected to the prokaryotic expression vector pET30a and transformed into Escherichia coli BL21 to induce expression.After the detection of SDS-PAGE,bands of target protein were identified.It was further confirmed that there was a difference in SiIND1 protein expression size between the capsule dehiscent variety and indehiscent variety,and there was a premature termination of translation in capsule indehiscent variety.

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