Genetic Diversity and Molecular Evolution of Chinese Waxy Maize Germplasm

Waxy maize (Zea mays L. var. certaina Kulesh), with many excellent characters in terms of starch composition and economic value, has grown in China for a long history and its production has increased dramatically in recent decades. However, the evolution and origin of waxy maize still remains unclear. We studied the genetic diversity of Chinese waxy maize including typical landraces and inbred lines by SSR analysis and the results showed a wide genetic diversity in the Chinese waxy maize germplasm. We analyzed the origin and evolution of waxy maize by sequencing 108 samples, and downloading 52 sequences from GenBank for the waxy locus in a number of accessions from genus Zea. A sharp reduction of nucleotide diversity and significant neutrality tests (Tajima’s D and Fu and Li’s F*) were observed at the waxy locus in Chinese waxy maize but not in nonglutinous maize. Phylogenetic analysis indicated that Chinese waxy maize originated from the cultivated flint maize and most of the modern waxy maize inbred lines showed a distinct independent origin and evolution process compared with the germplasm from Southwest China. The results indicated that an agronomic trait can be quickly improved to meet production demand by selection.     

Identification of Genetic Differentiation between Waxy and Common Maize by SNP Genotyping

Waxy maize (Zea mays L. var. ceratina) is an important vegetable and economic crop that is thought to have originated from cultivated flint maize and most recently underwent divergence from common maize. In this study, a total of 110 waxy and 110 common maize inbred lines were genotyped with 3072 SNPs to evaluate the genetic diversity, population structure, and linkage disequilibrium decay as well as identify putative loci that are under positive selection. The results revealed abundant genetic diversity in the studied panel and that genetic diversity was much higher in common than in waxy maize germplasms. Principal coordinate analysis and neighbor-joining cluster analysis consistently classified the 220 accessions into two major groups and a mixed group with mixed ancestry. Subpopulation structure in both waxy and common maize sets were associated with the germplasm origin and corresponding heterotic groups. The LD decay distance (1500–2000 kb) in waxy maize was lower than that in common maize. Fourteen candidate loci were identified as under positive selection between waxy and common maize at the 99% confidence level. The information from this study can assist waxy maize breeders by enhancing parental line selection and breeding program design.     

Molecular characteristics of two new <Emphasis Type="Italic">waxy</Emphasis> mutations in China waxy maize

The waxy gene mutation causes waxy maize grain to have a sticky quality. China has numerous waxy maize landraces and is thought to be the place of origin of waxy maize. The most abundant waxy maize resources in China are located in the Yunnan province and its surrounding areas. We collected 57 waxy maize landraces from Yunnan province and cloned and sequenced the waxy gene from its fourth to eighth exon. Two new waxy gene mutations, named wx-Cin4 and wx-124, were identified. The wx-Cin4 mutation is a 466-bp retrotransposon inserted into exon six. The wx-124 mutation is a 116-bp miniature inverted-repeat transposable element inserted into exon seven. This is the first time a 124-type mutation has been found in a maize waxy gene. The discovery of the two specific waxy mutations from landraces collected in Yunnan province provides new evidence supporting the hypothesis that China is the origin area for waxy maize.     

7个品种变叶木过氧化物酶同工酶分析

本试验对7个品种变叶木的形态特征进行比较,并利用聚丙烯酰胺凝胶电泳,对各品种过氧化物酶同工酶进行分析。结果表明,各品种的过氧化物酶同工酶谱不一;7个品种变叶木共有14条谱带,2号谱带最多为11条,5号谱带最少为6条;其谱带分为3个区(A=0~0.18,B=0.18~0.35,C=0.35~0.5);并对7个品种的亲缘关系进行聚类分析。     

菊花不同生长阶段不同器官POD和EST同工酶比较

采用过氧化物酶(POD)、酯酶(EST)2个酶系统的12个同工酶位点,分析了4个菊花品种营养生长和生殖生长阶段不同器官(嫩叶、老叶、嫩茎、木质化茎)的同工酶变化.结果表明:(1)4个品种共有16种POD酶带,15种EST酶带;(2)菊花的POD和EST具有组织特异性和阶段特异性,其中以嫩叶的酶带最多,其次为老叶,再次为嫩茎,而木质化茎的酶带最少;(3)与生殖生长阶段相比,营养生长阶段的POD酶带更清晰,更整齐,分离更好,但生殖生长阶段的EST同工酶比营养生长阶段的更清晰;(4)营养生长阶段的嫩叶最适合用于菊花POD同工酶分析,而EST同工酶研究宜取生殖生长时期的嫩叶.     

The Amino Acid Sequence of Ascorbate Peroxidase from Tea Has a High Degree of Homology to That of Cytochrome c Peroxidase from Yeast

The chloroplast isozyme of ascorbate peroxidase from tea leaveswas digested with lysyl endopeptidase, and the amino acid sequencesof the peptide fragments were determined. These sequences accountedfor 64% of the amino acids in the entire protein. The sequenceof one of the peptides can be aligned with the region whichincludes the proximal histidine that serves as the fifth ligandof the heme iron in guaiacol peroxidases and cytochrome c peroxidase.The sequences of the peptides from ascorbate peroxidase exhibita higher degree of homology to the sequence of cytochrome cperoxidase from yeast than to those of guaiacol peroxidasesfrom plants. In addition, three of the peptides from ascorbateperoxidase show a high degree of homology to triose-phosphateisomerase from maize. From the available amino acid sequencesand the enzymatic and molecular properties of ascorbate andcytochrome c peroxidases, we propose that these hydrogen peroxide-scavengingperoxidases that use either cytochrome c or ascorbate as theelectron donor originated from the same ancestral protein. (Received July 5, 1991; Accepted December 6, 1991)     

The Role of Peroxidase and Polyphenol Oxidase Isozymes in Wheat Resistance to Alternaria triticina

Polyphenol oxidase activity was higher in resistant wheat cultivar ACC-8226 than in susceptible cultivar MP-845 in control sets and after inoculation of Alternaria triticina. However, similar polyphenol oxidase isozyme pattern was found in control and inoculated sets of both the cultivars, but the band intensity was higher after inoculation. Three and four peroxidase isozymes were found in ACC-8226 and MP-845, respectively. An extra peroxidase isozyme band was observed in both the cultivars after inoculation. The results suggest an active role of peroxidase and polyphenol oxidase in defence mechanism of wheat plants.     

湖北大麦品种酯酶同工酶酶谱与地区分布关系的研究

对产于湖北的１０５份大麦品种,采用聚丙烯酰胺凝胶电泳进行了酯酶同工酶酶谱分析,共得到８种酶谱类型。８种酶谱聚为４类,其中具酶谱类型Ｉ、Ⅱ、Ⅲ和Ⅷ的品种约占供试材料的９４％,构成湖北大麦酯酶同工酶酶谱的主要类型。对这些品种来源地的分析表明,具有相同或相近酶谱的品种分布在一些相对集中的地区,这些地区的气候条件较相似。对湖北大麦酯酶同工酶酶谱类型与地区分布和气温关系的关系,以及酯酶在大麦引种育种生产实践     

POD同工酶在酸枣、枣分类中的应用

运用聚丙烯酰胺凝胶电泳（ＰＡＧＥ）技术,对来自不同产地的８３个酸枣类型（品种）,及３６个枣品种的叶片过氧化物酶（ＰＯＤ）同工酶扫描数据,进行了主因子分析和聚类分析研究。结果表明,运用距离系统的类平均法（ＵＰＧＭＡ）可将全部试材划分为六大类群和３个特殊型,且聚类结果与地理位置直接相关。此外,依同工酶带的有无,可将全部供试枣品种叶片ＰＯＤ同工酶合并为５种谱型,全部酸枣类型（品种）合并为２２种谱型。大马     

应用同工酶进行宽皮柑桔分类及其进化研究

利用聚丙烯酰胺凝胶电泳法,分析了50个宽皮柑桔生物型及5个甜橙品种的过氧化物酶、谷氨酸草酰乙酸转氨酶、四唑氧化酶、6-磷酸葡萄糖异构酶、6-磷酸葡萄糖变位酶、超氧化物歧化酶、苹果酸酶以及酯酶等8种酶系统同工酶。运用数量分类学的原理及方法,对同工酶资料进行了聚类分析。结果表明,宽皮柑桔的起源是多元的,小果类桔属于宽皮柑桔的原始类型,在进化过程中起着较为重要的作用。中国原产宽皮柑桔的代表种以道县野桔、马鼻桔或狗屎柑为佳。柑的来源有两条途径,即桔橙杂种和桔的单元演化;蕉柑是栟柑与甜橙的杂种;温州蜜柑来源于黄岩本地广桔的实生变异。     

Analysis of genetic diversity and relationships among waxy maize inbred lines in Korea using SSR markers

Information regarding the genetic diversity and genetic relationships among elite inbred lines is necessary to improve new cultivars in maize breeding programs. In this study, genetic diversity and genetic relationships were investigated among 84 waxy maize inbred lines using 50 SSR markers. A total of 269 alleles were identified at all the loci with an average of 5.38 and a range between 2 and 13 alleles per locus. The gene diversity values varied from 0.383 to 0.923 with an average of 0.641. The cluster tree generated using the described SSR markers recognized two major groups at 32% genetic similarity. Group I included 33 inbred lines while group II included 51 inbred lines. The clustering patterns of most of the waxy maize inbred lines did not clearly agree with their source, pedigree or geographic location. The average GS among all inbred lines was 35.7 ± 10.8. Analysis of waxy maize inbred lines collected from Korea and China at 50 SSR loci revealed higher values of average number of alleles (4.9) and gene diversity (0.638) in Korean inbred lines as compared to Chinese inbred lines (3.5 and 0.563, respectively). The information obtained from the present studies would be very useful for maize breeding programs in Korea.     

多花水仙若干品种类型的亲缘关系与进化研究I.POD同工酶分析

以7个水仙品种为材料,用垂直平板聚丙烯酰胺凝胶电泳,分析在同一生长时期的过氧化物酶同工酶。结果表明,酶谱中不同品种的酶带分布有区别。但I-1、I-2和Ⅲ-3相似,Ⅱ-1和Ⅱ-2相似,平潭水仙和漳州水仙相似。     

ISSR analysis of genetic diversity in sacred lotus cultivars

In this study, inter-simple sequence repeats (ISSR) markers were applied to assess genetic diversity and genetic relationships of 92 cultivars of sacred lotus (Nelumbo nucifera Gaertn.), one of the most famous flowers in China. Our results showed that sacred lotus exhibited a low level of genetic diversity (percentage of polymorphic bands, PPB = 55.8%), which may result from its asexual mode of reproduction and long-term artificial selection. Clustering analyses indicated that these cultivars could be divided into two clades. Most cultivars of Chinese lotus species origin were included in one clade, and one cultivar of American lotus species origin was nested in the other clade. The hybrid cultivars from hybridization between the two subspecies were interspersed in these two clades. Seven cultivars native to Thailand formed a distinct subclade among the cultivars of Chinese lotus species origin. Genetic differentiation between two subspecies, and between cultivars from Thailand and other cultivars could be attributed to geographic isolation. The monophyly of three cultivars of Sanshui Winter Lotus and their closest relationships to Chinese lotus species origin suggests that they might have a common origin and may consist completely or mainly of genetic material from N. nucifera subsp. nucifera.     

Preliminary Studies on the Relationship between Diterpene Quinone Compounds and Peroxidase Isozyme in the Labiatae

Peroxidase isozyme was determined by polyacrylamide gel column electrophoresis method in 18 species of the Labiatae. Results showed that the band of peroxidase isozyme of those species containing diterpene quinone compounds appeared in the middle of the column with an inclination to the anode. The contents of diterpene quinone compounds in the plant were usually consistant with the band of its peroxidase isozyme. The band of peroxidase isozyme of those species containing essential oil almost did not appear in the column under the same condition.     

Different maternal origins of Japanese lowland and upland rice populations

Plastid subtype ID (PS-ID) sequences were determined from sequence data based on CA repeats between genes rpl16 and rpl14 in Japanese lowland and upland cultivars. The PS-ID sequences of Japanese rice cultivars showed that there are different maternal origins between lowland and upland cultivars. One subtype, 6C7A, of PS-ID sequences was predominant in all but one Japanese lowland cultivar and carried a combination of the indica-specific subtype 8C8A and japonica-specific nuclear markers for the isozyme genotype. It is probably a nuclear-cytoplasmic recombinant resulting from natural out-crossing and succeeding self-pollination. The origin of the plastid was re-confirmed by the existence of an indica-specific deletion in the plastid genome. In contrast, the Japanese upland cultivars showed two subtypes, 7C6A and 6C7A, of PS-ID sequences. An upland-specific isozyme allele as a nuclear marker was equally predominant in cultivars carrying each subtype. The existence of these particular upland-specific nuclear and cytoplasmic genotypes suggests that the origin of Japanese upland cultivars is different from that of Japanese lowland cultivars. Cultivars carrying the upland-specific nuclear genotype are common in Southeast Asia, but the combination of the upland-specific nuclear and cytoplasmic genotypes which is the same as the Japanese upland predominant type was found in cultivars only in Taiwan and Indonesia. Japanese upland cultivars are closely related to those cultivars.     

玉米酯酶同工酶和过氧化物酶同工酶的分子量研究

本试验利用聚丙烯酰胺凝胶梯度电泳分步染色法直接对玉米苗期酯酶同工酶和过氧化物酶同工酶各酶带的分子量进行了比较测定。酯酶同工酶 E_1、E_2、E_3~F、E_3~S、a、b、c 各酶带的分子量分别为<20000,35200、33000、38500、29900、28500、34000道尔顿过氧化物酶同工酶 PX_4~F和 PX_4~S酶带的分子量分别为131000和149000道尔顿。根据酶带在均匀胶和梯度胶中的位置变化对各酶带的生化性质作了初步分析,发现 E_3~F和 E_3~S、PX_4~F 和 PX_4~S 在迁移率上的差异主要是分子量的差异。本文为同工酶的分子量测定提供了一个简便的方法。     

Studies on the Molecular Weights of Esterase and Peroxidase Isozymes of Maize

The molecular weights of esterase and peroxidase isozymes of maize seedlings were directly determined by improved polyacrylamide gradient gel electrophoresis. The different isozyme bands developed in polyacrylamide slab gel electrophoresis (uniform gel) were identified in polyacrylamide gradient gel electrophoresis by means of isozyme variants. The molecular weights of esterase isozymes E1, E2, E3F, E3S, a, b, c, named according to isozyme patterns in uniform gel, are <20000, 35200, 33000, 38500, 29900, 28500, 34000 doltons respectively. The molecular weights of peroxidase isozymes PX4F and PX4S are 131000 and 149000 doltons respectively. According to the band location in uniform gel and in gradient gel, some biochemical properties of the isozyme bands and relationships between the isozyme bands were analyzed. The possible errors in the determination of smaller molecular weight isozymes are discussed.     

Variability in antifungal proteins in the grains of maize, sorghum and wheat

Crude protein extracts were made from kernels of 12 cultivars each of maize, sorghum and wheat. These preparations were fractionated on sodium dodecyl sulfate (SDS)-polyacrylamide gels and subjected to Western blot analyses. Bands corresponding to chitinases and β-glucanases were identified immunologically (Western blots) and on activity gels. Ribosome Inactivating Protein(s) (RIP) and permatins were identified immunologically. In maize, two chitinase bands (25–29 kDa) were seen in all cultivars, and a third band of about 23 kDa was detected in 7 of the 12 cultivars. Two or three β-glucanase bands of sizes between 24 and 36 kDa (probably a mixture of 1,3–β- and 1,3–1,4-β-glucanases) were detected in blots of SDS gels, and one band was detected in activity gels (1,3-β-glucanase). In sorghum, one chitnase band of approximately 29 kDa, and two or three additional bands ranging in size from 21–24 kDa were observed. Only one β-glucanase band was identified, with an estimated molecular weight of 30 kDa. All bands that appeared on Western blots of SDS gels corresponded to bands detected on activity gels. In wheat, one chitmase band of around 20 kDa, one β-glucanase band of about 30 kDa and one RIP band of about 30 kDa were identified. Permatins (molecular weight about 22 kDa) were identified in maize, sorghum and wheat, with the different cultivars having varying amounts of permatins.     

He-Ne激光对啤酒大麦麦芽同工酶基因表达的影响

采用不同时间He-Ne激光（5 m W/mm2）对＂临啤二号＂大麦种子进行辐照,利用聚丙烯酰胺凝胶电泳（PAGE）技术分析激光辐照对大麦麦芽中超氧化物歧化酶（SOD）同工酶、过氧化物（POD）同工酶、过氧化氢（CAT）同工酶、β-淀粉酶同工酶、三磷酸腺苷（ATPase）同工酶酶谱的影响。结果表明：种子经He-Ne激光辐照后诱导出1条新POD同工酶谱带,且总POD活性在60 s和90 s处有所增加;30 s激光辐照诱导出1条新的CAT同工酶谱带,其它时间处理酶的组分没有变化,但酶活性增加;He-Ne激光辐照使SOD、β-淀粉酶及ATPase同工酶的组分在不同时间下有所减少,但一定时间的激光辐照提高了其同工酶的活性。综合实验结果可知,60 s～90 s He-Ne激光可以促进大麦种子萌发过程中以上同工酶基因的表达,有利于缩短制麦周期,提高麦芽糖化力及抗氧化力。