Scientia Agricultura Sinica ›› 2023, Vol. 56 ›› Issue (10): 1838-1847.doi: 10.3864/j.issn.0578-1752.2023.10.002

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Molecular Marker Assisted Identification and Application of Maize Germplasms for Maize Rough Dwarf Disease Resistance

WANG JiangHao(), WANG LiWei, ZHANG DongMin, GUO Rui, ZHANG QuanGuo, LI XingHua, WEI JianFeng, SONG Wei, WANG BaoQiang(), LI RongGai()   

  1. Institute of Cereal and Oil Crops, Hebei Academy of Agriculture and Forestry Sciences/Hebei Laboratory of Crop Genetics and Breeding, Shijiazhuang 050035
  • Received:2022-12-17 Accepted:2023-02-13 Online:2023-05-16 Published:2023-05-17

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Abstract:

【Objective】Molecular markers tightly linked to three maize rough dwarf disease (MRDD) resistant loci were employed to identify resistant inbred lines, then the classification of heterotic groups and analysis of combining ability of these inbred lines were carried out, which proved a highly efficient way for maize MRDD resistance breeding.【Method】A recombinant inbred lines (RILs) population consisting of 263 F9 lines was developed through single seed descent method from a segregating F2 population by crossing a resistant inbred line K36 to a susceptible inbred line S221. The MRDD resistances of the RILs were identified in different growing environments. Meanwhile the RILs were genotyped by employing three pairs of molecular markers, 5FR, 6W53 and IDP25K which were closely linked to the three resistant loci, qMrdd2, Rmrdd6 and qMrdd8. The excellent lines with disease resistance and good agronomic traits were selected out after field evaluation. Totally 24 maize inbred lines including the elite lines were genotyped using Maize 56K SNP array, then the genetic distances between the selected lines and other elite inbred lines were calculated according to Roger's algorithm and cluster analysis was conducted to classify the heterotic groups. Meanwhile, hybrid combinations were generated and the combining abilities were tested to screen the combinations with strong disease resistance and heterosis.【Result】The inbred line K36 were homozygous resistant at the three loci, qMrdd2, Rmrdd6 and qMrdd8 while S221 were homozygous susceptible. All the 263 RILs were genotyped into 21 patterns in terms of genetic composition of the three resistant loci. The lowest DSI (0.281) appeared when all the three loci were homozygous resistant while the highest DSI (0.776) appeared when the three loci were homozygous susceptible, which were consistent with the resistant and susceptible parents (0.257, 0.623). The order of DSI from low to high value for one homozygous resistant locus was Rmrdd6 (0.396), qMrdd8 (0.478) and qMrdd2 (0.654) when the other two loci were homozygous susceptible, which showed that Rmrdd6 and qMrdd2 performed the strongest and the weakest resistance while qMrdd8 was in the middle. The variation range of genetic distance between JR2136 with the genotype of three homozygous resistant loci and other 23 inbred lines was 0.2234-0.2895, with an average value of 0.2612. The inbred line with the smallest genetic distance was C413, and the largest was Chang7-2. According to the results of cluster analysis, JR2136 was classified into Reid group, hybrid combinations with inbred lines H92 and H521 belonging to Huanggai group performed strong disease resistance and heterosis.【Conclusion】The resistance of K36 to MRDD was controlled by three loci, qMrdd2, Rmrdd6 and qMrdd8, and it had quantitative genetic characteristics and gene additive effect. Maize varieties with homozygous resistant genotypes demonstrated the strongest disease resistance. The developed molecular markers closely linked with the three resistant loci have proved valuable tools in disease-resistant breeding and screening of resistant germplasm resources. It is feasible to use molecular markers for assisted selection and gene aggregation to select highly heterotic combinations with strong disease resistance.

Key words: maize rough dwarf disease, resistance gene, molecular marker-assisted selection (MAS), heterotic grouping, combining ability

Table 1

Disease resistance identification results of 263 plant lines in RILs"

抗性等级 Resistance level 地点Site 免疫Immune 高抗HR 抗R 感S 高感HS
病情指数DSI 0 0.00—0.33 0.34—0.66 0.67—0.99 ≥1.00
病家系数量
No. of diseased lines		 河北藁城 Gaocheng, Hebei 0 43.0 192.0 26.0 2.0
山东济宁Jining, Shandong 0 28.0 105.0 77.0 53.0
病家系率
Rate of diseased lines (%)		 河北藁城Gaocheng, Hebei 0 16.3 73.0 9.9 0.8
山东济宁Jining, Shandong 0 10.8 40.0 29.2 20.0

Table 2

Comparison of maize rough dwarf disease incidence in Shandong and Hebei in 2019"

调查地点
Sites		 播种期
Sowing date (M/D)		 病情指数DSI
RILs K36 S221
河北藁城 Gaocheng, Hebei 5/18 0.21 0.02 0.54
山东济宁 Jining, Shandong 5/16 0.49 0.28 0.62

Fig. 1

DSI distribution of RILs population in Hebei and Shandong environments"

Fig. 2

Genotype of inbred lines and RILs population families at the resistant sites on chromosomes 2, 6 and 8 of maize A：IDP25K；B：MGS；C：5FR；D：6W53；E：6W19-95；1—8：100 bp DNA marker、S221、K36、JR2136、B73、X178、Ye478、Qi319。F：IDP25K；G：5FR；H：6W53；1—39：50 bp DNA marker、C1—C36、S221、K36"

Table 3

DSI of different genotype plant lines with three resistance loci in RILs population"

材料名称
Name of
varieties		 家系抗性位点基因型
Genotype of resistant locus in plant lines		 平均病情指数
DSI
qMrdd2 Rmrdd6 qMrdd8
K36 R R R 0.257
S221 S S S 0.623
RILs家系
RILs		 R R R 0.281
S S S 0.776
S R R 0.454
R S R 0.622
S R S 0.396
S S R 0.478
R S S 0.654
S S H 0.799
S H S 0.475
H S S 0.625
R H R 0.313
H R R 0.313
R R H 0.469
S H R 0.503
H H R 0.521
R H S 0.515
H R S 0.30
R S H 0.653
R H H 0.477
S H H 0.447
H H H 0.438

Fig. 3

UPGMA tree for 24 inbred lines based on Roger’s genetic distance"

Table 4

Comparison of important agronomic traits and yield traits of hybrid combinations"

杂交种
Hybrids		 产量
Yield
(kg·hm-2)		 与对照相比
Compared with control (%)		 生育期
Growth period (d)		 株高
Plant height (m)		 穗位
Ear height (m)		 穗长
Ear length (cm)		 穗行数
No. of rows		 行粒数
No. of grains per row		 百粒重
Weight per 100 grains (g)
JR2136×H92 11334.90 7.01 106 2.95 1.29 19.0 14.8 41.4 34.5
JR2136×H521 11418.75 7.80 107 2.95 1.31 20.0 14.8 42.0 36.8
郑单958 Zhengdan 958 (CK) 10592.40 0.00 106 2.92 1.28 14.4 14.6 32.2 34.7
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