Crops ›› 2024, Vol. 40 ›› Issue (5): 1-7.doi: 10.16035/j.issn.1001-7283.2024.05.001

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Phenotypic Analysis and Gene Mapping of Rice Mutant Chalkiness and Shrunken Endosperm-2

Sun Jiameng(), Gao Yuan, Chen Hu, Hua Qin, Lin Quanxiang, Chen Qingquan, Li Jincai, Zhang Haitao()   

  1. College of Agronomy, Anhui Agricultural University, Hefei 230036, Anhui, China
  • Received:2023-06-24 Revised:2023-08-12 Online:2024-10-15 Published:2024-10-16

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Abstract:

Starch is the most abundant macromolecular substance in rice grain, and its composition and content have direct effect on quality indexes of rice. In this paper, agronomic and qualitative traits related to chalkiness and shrunken endosperm-2 (cse-2) were investigated and the candidate gene was mapped. The results showed that the agronomic traits including plant height, grain number per panicle, seed-setting rate, starch content and its components, physicochemical properties, starch grain morphology and structure, protein and fatty acid contents of the mutant had significantly altered. Mapping result showed that mutated phenotype is determined by a pair of nuclear recessive genes locating on chromosome 4. Sequence analysis indicated that a single nucleotide substitution of C-to-T occurred on the 6064th base of LOC_Os04g55230/FLO2, forming a TAA nonsense mutation. Therefore, cse-2 was a new allelic mutant of flo2.

Key words: Rice, Floury endosperm, Starch property, Gene mapping, FLO2

Fig.1

Comparison of phenotypes between wild type (ZH11) and mutant (cse-2) Plant height (a) and grain (b-e). Bars=10 cm (a), 1 cm (b-d) and1 mm (e)."

Table 1

Comparison of agronomic traits between wild type (ZH11) and mutant (cse-2)"

农艺性状Agronomic trait ZH11 cse-2
株高Plant height (cm) 104.05±0.56 94.55±1.18**
主茎穗长Main panicle length (cm) 23.80±0.46 21.22±0.51**
第1节间长
The first internode length (cm)		 42.20±0.66
 40.21±0.75
第2节间长
The second internode length (cm)		 19.16±0.64
 18.57±0.40
第3节间长
The third internode length (cm)		 13.68±0.56
 10.55±0.56**
第4节间长
The fourth internode length (cm)		 7.35±0.14
 5.24±0.17**
有效分蘖数Effective tiller number 20.75±2.47 24.85±2.56
一次枝梗数Primary branch number 13.88±0.40 13.83±1.01
二次枝梗数Secondary branch number 44.00±1.40 35.50±1.76**
穗粒数Grain number per panicle 221.85±6.84 191.90±6.11**
结实率Seed-setting rate (%) 89.84±0.13 80.58±0.19**
粒长Grain length (mm) 7.68±0.06 7.67±0.09
粒宽Grain width (mm) 3.51±0.01 3.49±0.02
粒厚Grain thickness (mm) 2.35±0.03 2.04±0.02**
千粒重1000-kernel weight (g) 25.63±0.40 19.16±0.17**

Fig.2

SEM images of grain cross section of wild type (ZH11) and mutant (cse-2)"

Fig.3

Comparison of grain component contents of wild type (ZH11) and mutant (cse-2) “**”indicates extremely significant difference at P < 0.01 level."

Table 2

Comparison of the rice phypical and chemical properties between wild type (ZH11) and mutant (cse-2)"

材料
Material		 起始温度
To (℃)		 峰值温度
TP (℃)		 结束温度
TC (℃)		 焓变
?H (J/g)		 碱消值
Alkali consumption value		 胶稠度
Gel consistency (mm)
ZH11 66.90±0.55 72.20±0.37 76.10±0.27 6.50±0.22 8.10±0.37 56.40±0.18
cse-2 62.20±0.31** 68.10±0.48** 73.60±0.17** 5.80±0.11** 7.70±0.48** 69.60±0.15**

Fig.4

Fine mapping of CSE-2 gene a-d: the numbers above and beneath the bold lines represent the molecular markers and recombinants, respectively, and the light blue arrow represent the candidate gene; e: the structure diagram of candidate gene CSE-2, the light blue box represent the exon, the blank part represent the intron, the red arrows represent the start codon, stop codon and mutation site, respectively, and the content in the black box represent the mutation; f: target gene reverse sequencing peak map, with mutated sites in the red line box; g: partial protein sequence alignment near the none-sense mutation site."

Fig.5

Sequence alignment of FLO2 protein between wild type (ZH11) and mutant (cse-2) Red arrow indicates the site that the stop codon occurred of FLO2 in the cse-2 mutant."

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