摘要
本研究以山东农业大学选育的6个“丰花”系列花生新品种为主要材料,采用SSR分子标记技术研究该系列品种与其亲本以及其它栽培品种(44个)间的DNA差异,筛选获得品种的特异标记,并分析特异标记的遗传来源和在其它栽培品种中的出现几率,以评价“丰花”系列花生品种所聚合亲本遗传物质的特点。选用4个“丰花”系列品种与其它4个栽培品种按GriffingⅡ配制杂交组合,考察亲本和F1代植株叶片的光合速率、叶绿素含量、SOD活性、POD活性和MDA含量,对上述生理性状进行遗传分析。主要结果如下:
1. 59个花生品种SSR指纹图谱及遗传特异性分析
利用80对SSR引物进行59个品种的PCR扩增,筛选出7对多态性引物,共扩增出78个条带,其中72条具有多态性,多态性频率为92.31%。构建了59个栽培品种的SSR指纹图谱。有13个品种只能通过二岐分类法与其它品种鉴别开。丰花6号可以由引物P59进行PCR扩增鉴别;丰花1和丰花5分别可以用引物P5和引物P48进行鉴别;丰花3号可以用引物P5、引物P47和引物P48进行鉴别;丰花2号可以由引物P12、引物P44、引物P59及引物P74进行鉴别。
2.“丰花”系列品种和亲本间的DNA差异分析
丰花1号的29个多态性标记位点中,来自母本(蓬莱一窝猴)的占3.4%,来自父本(海花1号)的占65.5%,不同于亲本的特异标记位点占31.03%;丰花2号、丰花3、丰花4号、丰花5号分别能检测到1、3、3、7个与双亲均不同的特异位点。表明育成的“丰花”系列品种遗传来源不是父、母本的简单组合,辐射诱变丰富了其遗传物质基础。
3.“丰花”系列品种中双亲SSR标记位点在其它品种中的分布
分析“丰花”系列品种来自父本或母本的SSR标记位点在其它主栽品种(共44个)中出现的几率的结果表明:丰花1号来自父本的标记位点P5G的几率为22.7%;丰花2号来自父本的标记位点P74A和P74E的几率仅为6.8%;丰花4号来自母本的标记位点P59J的几率为20.5%,来自父本的标记位点P28E的几率为19.2%;丰花6号来自母本的标记位点P76B的几率为22.7%,来自父本的标记位点P33A的几率为24%。表明“丰花”系列品种聚合了亲本的稀有基因源。
4.叶片生理性状配合力效应分析
比较8个品种间各生理性状的一般配合力表明:A596光合速率的一般配合力较高,效应值为1.38,丰花5号的叶绿素含量的一般配合力较高,效应值为0.11,丰花2号SOD活性的一般配合力较高,效应值为86.60,荔浦大花生POD活性的一般配合力较高,效应值为12.51,MDA含量配合力方差差异不显著。
比较28个杂交组合的特殊配合力的结果表明:光合速率特殊配合力较高的组合有丰花5号/荔浦大花生、白沙1016/丰花5号和丰花2号/丰花1号,其效应值依次为:2.89、1.49和1.43;丰花2号/白沙1016组合叶绿素含量的特殊配合力最高,效应值为0.29;SOD活性特殊配合力较高的组合有丰花2号/白沙1016和丰花1号/鲁花11,效应值分别为286.52和224.51;POD活性特殊配合力最高的组合为丰花5号/荔浦大花生,其次为丰花3号/荔浦大花生,效应值分别为20.21和6.62;A596/荔浦大花生MDA含量的特殊配合力有最高的负向效应,效应值为-0.60。
5.花生叶片生理性状的遗传模型
光合速率、SOD活性、POD活性及MDA含量四个生理性状的Wr-Vr的方差均达显著或极显著水平,表明这些性状的遗传受上位性效应的影响,遗传均符合“加性—显性—上位性”模型。
6.花生叶片生理性状的遗传力估算
SOD活性和POD活性的广义遗传力分别为94.99%和97.46%,狭义遗传力分别为63.83%和54.35%;MDA含量的狭义遗传力仅为0.50%,广义遗传力是93.34%;光合速率与叶绿素含量的狭义遗传力分别为为34.70%和28.85%,广义遗传力分别是84.38%和90.10%。SOD活性、POD活性、光合速率和叶绿素含量均可在早代选择,MDA含量适宜晚期世代选择。
7.花生叶片生理性状的杂种优势评价
在所有杂交组合中,丰花2号/丰花1号各生理性状除POD活性表现出负向的超亲优势(-21.26%)和平均优势(-11.63%)外,其余性状均表现出较高的杂种优势,其中叶绿素含量的杂种优势表现最明显,超亲优势为25.0%,平均优势为27.12%。
The six“Fenghua”peanut cultivars were main materials in this research. Specific molecular markers of“Fenghua”cultivars were found out through studying the difference between“Fenghua”cultivars and their parents and the difference between“Fenghua”cultivars and other 44 peanut varieties by SSR. The characteristic aggregation genetic material from the parents of“Fenghua”can be evaluated through the analysing genetic origins of specific markers and the odds in other varieties; four varieties of Fenghua cultivars and other four cultivars varieties were used to confect cross combination by GriffingⅡ. Investigated Photosynthesis rate, chlorophyll content, SOD activities, POD activities and MDA content of F1 and their parents and studied variance, combining ability, genetic model, heritability and heterosis of these physiological characters be investigated. .The results were summarized as follow.
1. SSR fingerprint of 59 peanut varieties and analysis of their genetic specificity
Seven pairs of SSR primers out of eighty identified have polymorphism bands when PCR of 59 varieties that 78 bands in total land were produced and 72 of whom were polylmorphic bands, accounted for 92.31%. SSR fingerprint was constructed of 59 peanut cultivars. 13 cultivars should be identificated with other cultivars by Second-chimata-classification. Fenghua 6 can be identificated by primer P59; Fenghua 1 and Fenghua 5 can be identificated by primer P5 and primer P48; Fenghua 3 can be identificated by primer P5, P47 and P48; Fenghua 2 can be identificated by primer P12, P44, P59 and P74.
2. Analysis of DNA difference between Feng-hua and their parents
In all 29 loci polymorphic markers of Fenghua 1,the markers from his mother (Penglaiyiwohou) was accounted for 3.4%, from his father (Haihua 1) accounted for 65.5% and the specificity different from the parent marker loci accounted for 31.03%; 1, 3, 3, 7 specific markers can be detected of Fenghua 2, Fenghua 3, Fenghua 4 and Fenghua 5 with their parents. Which showed that the genetic origin of Fenghua was not simple combination of male and female, radiation-induced mutation and strict selection of filial generation enrich the genetic basis more.
3. Distribution of SSR sites of Feng-hua root in parents on other 44 peanut varieties
Analysis of "Floribunda" series from their fayher or their mother species of SSR marker loci varieties grown in other (a total of 44) chance of appearing in the results showed that: the P5G site from father(Haihua 1) on Fenghua 1 accounted for 22.7%; the P74A and P74E sites from father(Lipudahuasheng) on Fenghua 2 accounted for 6.8%; the P59J site from mother(Baisha 1016) on Fenghua 4 accounted for 20.5%, the P28E site from father(A596), accounted for 19.2%; the P76B site from mother(Baisha 1016) on Fenghua 6 accounted for 22.7%, the P33A site from father(Burma peanut), accounted for 24%. Which showed that“Fenghua”converge far-between genome of their parents.
4. Combining ability effects analysis of physiological characters of leaves
Comparison of general combining ability of the physiological traits between the eight species showed that: A596 had higher general combining ability on photosynthetic rate, effect value of 1.38; Fenghua 5 had higher general combining ability on chlorophyll content with the effect value of 0.11; Floribunda on the 2nd general combining ability effect of SOD activity of Fenghua 2 was higher (86.60), Lipudahuasheng had higher general combining ability effect on POD activity in with 12.51, the combining ability variance MDA content had no obvious difference.
The results of Comparison of 28 cross combinations of specific combining ability showed that: Fenghua 5/Lipudahuasheng, Baisha 1016/Fenghua 5, Fenghua 2/Fenghua 1 had higher specific combining ability on photosynthetic rate, the effect were: 2.89,1.49 and 1.43; special combination on chlorophyll content of Fenghua 2/Baisha 1016 was the highest (0.29);Fenghua 2/Baisha 1016 and Fenghua 1/Luhua 11 had higher specific combining ability on SOD activity with the effect of 286.52 and 224.51; POD activity of specific combining ability of the combination of the highest on the Fenghua 5/Lipudahuasheng, followed by the Fenghua 3/Lipudahuasheng, effect values were 20.21 and 6.62; MDA content of A596/Lipudahuasheng in the highest specific combining ability effect of the negative, effects of a value of -0.60.
5. Genetic model of physiological characters of leaves on peanut The Wr-Vr variance of photosynthesis rate, SOD activity, POD activity and MDA content reached remarkable level or super remarkable level that they exist epistatic interactions effect and they fit the additive- dominance- epistasis model.
6. Heritability of physiological characters of leaves on peanut The broad-sense heritability SOD activity and POD activity were 97.46% and 94.99%, respectively, the narrow heritability of 63.83% and 54.35%; the narrow sense heritability of MDA content was only 0.50%, broad-sense heritability was 93.34%; the narrow heritability of photosynthetic rate and chlorophyll content, respectively for 28.85% and 34.70%, broad-sense heritability were 84.38% and 90.10%. SOD activity, POD activity, photosynthetic rate and chlorophyll content can be selected in the early generation, MDA content in appropriate choice of the late generations.
7. Evaluation of heterosis of physiological characters of leaves on peanut
In all cross combination, the physiological characters without POD activities of Fenghua 2/Fenghua 1 has negative over-parent heterosis (-21.26) and mid-parent heterosis that other characters all have large heterosis. The heterosis of chlorophyll content is most obvious that the over-parent heterosis is 25.0 and the mid-parent heterosis is 27.12. They also have obvious heterosis of some characters when“Fenghua”were used as parents to compound cross combination that the heterosis of these characters can be used in production.
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