Rice Science ›› 2015, Vol. 22 ›› Issue (4): 171-179.DOI: 10.1016/S1672-6308(14)60295-X
收稿日期:2015-03-07
接受日期:2015-04-21
出版日期:2015-07-28
发布日期:2015-05-27
. [J]. Rice Science, 2015, 22(4): 171-179.
| M1 line | M2 line | Seed source | Mutagenesis | Phenotype of M3 | Single gene recessive mutation | Mutant designation |
| 6-1 | 6-1A | 2009 | 1 mmol/L NaN3 / 16 h presoak | Wet leaf/glossy segregating | Nd | wsl3 |
| 6-1 | 6-1B | 2009 | 1 mmol/L NaN3 / 16 h presoak | Wet leaf/glossy | Nd | |
| 7-17 | 7-17A | 2009 | 1 mmol/L NaN3 / 16 h presoak | Wet leaf/glossy segregating | Yes | wsl4 |
| 11-39 | 11-39A | 2009 | 1 mmol/L NaN3 / 20 h presoak | Wet leaf/glossy | Nd | wsl5 |
| 26 | 26.1 | 2011 | 2 mmol/L NaN3 / 16-20 h presoak | Wet leaf/glossy segregating | Yes | wsl6 |
| 264 | 264.2 | 2011 | 2 mmol/L NaN3 / 16-20 h presoak | Wet leaf/glossy segregating | Nd | wsl7 |
| 524 | 524.2 | 2011 | 2 mmol/L NaN3 / 16-20 h presoak | Wet leaf/glossy segregating | Yes | wsl8 |
| 680 | 680.2 | 2011 | 2 mmol/L NaN3 / 16-20 h presoak | Wet leaf/glossy segregating | Yes | wsl9 |
| 843 | 843.1 | 2011 | 2 mmol/L NaN3 / 16-20 h presoak | Wet leaf/glossy segregating | Yes | wsl10 |
| 1064 | 1064.2 | 2011 | 2 mmol/L NaN3 / 16-20 h presoak | Wet leaf/glossy | Nd | wsl11 |
| 1086 | 1086.2 | 2011 | 2 mmol/L NaN3 / 16-20 h presoak | Wet leaf/glossy | Nd | wsl12 |
| 1558 | 1558.1 | 2011 | 2 mmol/L NaN3 / 16-20 h presoak | Intermediate | Nd | |
| 1558 | 1558.2 | 2011 | 2 mmol/L NaN3 / 16-20 h presoak | Intermediate | Nd | wsl13 |
| Phenotypic ratios of segregating M3 populations were evaluated using Pearson’s Chi-square test for goodness-of-fit to the 3:1 (wild-type:mutant) ratio expected for single gene recessive mutations. Nd, Not determined; wsl, Wax crystal-sparse leaf. | ||||||
Table Genetic and physical characteristics of reduced epicuticular wax mutants.
| M1 line | M2 line | Seed source | Mutagenesis | Phenotype of M3 | Single gene recessive mutation | Mutant designation |
| 6-1 | 6-1A | 2009 | 1 mmol/L NaN3 / 16 h presoak | Wet leaf/glossy segregating | Nd | wsl3 |
| 6-1 | 6-1B | 2009 | 1 mmol/L NaN3 / 16 h presoak | Wet leaf/glossy | Nd | |
| 7-17 | 7-17A | 2009 | 1 mmol/L NaN3 / 16 h presoak | Wet leaf/glossy segregating | Yes | wsl4 |
| 11-39 | 11-39A | 2009 | 1 mmol/L NaN3 / 20 h presoak | Wet leaf/glossy | Nd | wsl5 |
| 26 | 26.1 | 2011 | 2 mmol/L NaN3 / 16-20 h presoak | Wet leaf/glossy segregating | Yes | wsl6 |
| 264 | 264.2 | 2011 | 2 mmol/L NaN3 / 16-20 h presoak | Wet leaf/glossy segregating | Nd | wsl7 |
| 524 | 524.2 | 2011 | 2 mmol/L NaN3 / 16-20 h presoak | Wet leaf/glossy segregating | Yes | wsl8 |
| 680 | 680.2 | 2011 | 2 mmol/L NaN3 / 16-20 h presoak | Wet leaf/glossy segregating | Yes | wsl9 |
| 843 | 843.1 | 2011 | 2 mmol/L NaN3 / 16-20 h presoak | Wet leaf/glossy segregating | Yes | wsl10 |
| 1064 | 1064.2 | 2011 | 2 mmol/L NaN3 / 16-20 h presoak | Wet leaf/glossy | Nd | wsl11 |
| 1086 | 1086.2 | 2011 | 2 mmol/L NaN3 / 16-20 h presoak | Wet leaf/glossy | Nd | wsl12 |
| 1558 | 1558.1 | 2011 | 2 mmol/L NaN3 / 16-20 h presoak | Intermediate | Nd | |
| 1558 | 1558.2 | 2011 | 2 mmol/L NaN3 / 16-20 h presoak | Intermediate | Nd | wsl13 |
| Phenotypic ratios of segregating M3 populations were evaluated using Pearson’s Chi-square test for goodness-of-fit to the 3:1 (wild-type:mutant) ratio expected for single gene recessive mutations. Nd, Not determined; wsl, Wax crystal-sparse leaf. | ||||||
Fig. 1. Wet leaf/glossy and intermediate-type mutant phenotypes. Leaves were subjected to a fine misting with water from a spray bottle. Water droplets adhere to the leaves of mutants and coalesce resulting in a wet, glossy appearance. A, Comparison of wild-type Sabine and the three mutants 6-1A, 7-17A, and 11-39A. Booting-stage flag leaves are shown; B, Comparison of the wild-type Sabine and the mutants 1558.2 and 6-1A. Fifth fully-expanded leaves from the base are shown. Unlike the other mutants, water droplets adhere to 1558.2 but do not coalesce and the leaves do not appear wet or glossy.
| Population | M3 seeds planted a | M3 plants phenotyped a | Segregation ratio (wild-type:mutant) | Pearson’s Chi-square test (3:1) | ||
| χ2 | P value | |||||
| 26.1 | 77 | 57 | 40:17 | 0.708 | 0.400 | |
| 680.2 | 77 | 61 | 43:18 | 0.661 | 0.416 | |
| 843.1 | 77 | 73 | 53:20 | 0.224 | 0.636 | |
| 524.2 | 95 | 68 | 53:15 | 0.314 | 0.575 | |
| a includes initial screening of M3 plants. | ||||||
Table 2 Segregation of wet leaf/glossyphenotype in four M3 populations.
| Population | M3 seeds planted a | M3 plants phenotyped a | Segregation ratio (wild-type:mutant) | Pearson’s Chi-square test (3:1) | ||
| χ2 | P value | |||||
| 26.1 | 77 | 57 | 40:17 | 0.708 | 0.400 | |
| 680.2 | 77 | 61 | 43:18 | 0.661 | 0.416 | |
| 843.1 | 77 | 73 | 53:20 | 0.224 | 0.636 | |
| 524.2 | 95 | 68 | 53:15 | 0.314 | 0.575 | |
| a includes initial screening of M3 plants. | ||||||
Fig. 2. Scanning electron micrographs of wild-type Sabine and 11 reduced epicuticular wax mutants.Adaxial leaf blade surface shown (knob-like structures are papillae). Scale bars, 10 μm.
| Line | Wax content (mg/g) | Reduction in wax content (%) |
| Sabine (wild-type) | 3.01 ± 0.49 | |
| 6-1A/B a | 0.91 ± 0.31 ** | 69.77 |
| 7-17A | 0.68 ± 0.42 ** | 77.41 |
| 11-39A | 1.38 ± 0.10 ** | 54.15 |
| Values are given as mean ± SD with four replicates except where noted. a, Due to insufficient tissue from 6-1A M3 mutants, additional samples were taken from 6-1B M3 mutants and content was determined using a total of five replicates (three from 6-1A plants, two from 6-1B plants included). ** means significant difference between mean values at P < 0.01 by t-test between the wild-type and each of the mutant line. | ||
Table 3 Total epicuticular wax content of wild-type and wsl mutant lines by weight method.
| Line | Wax content (mg/g) | Reduction in wax content (%) |
| Sabine (wild-type) | 3.01 ± 0.49 | |
| 6-1A/B a | 0.91 ± 0.31 ** | 69.77 |
| 7-17A | 0.68 ± 0.42 ** | 77.41 |
| 11-39A | 1.38 ± 0.10 ** | 54.15 |
| Values are given as mean ± SD with four replicates except where noted. a, Due to insufficient tissue from 6-1A M3 mutants, additional samples were taken from 6-1B M3 mutants and content was determined using a total of five replicates (three from 6-1A plants, two from 6-1B plants included). ** means significant difference between mean values at P < 0.01 by t-test between the wild-type and each of the mutant line. | ||
Fig. 3. Cuticle membrane permeability of three wax mutants. A, Water loss rate from detached leaves over time; B, Chlorophyll loss over time. Each bar represents the mean ± SD of two independent assays (two replicates for wild-type and four replicates for each mutant). Levels of significance between wild-type Sabine and each mutant were determined by t-test for unequal variance; * and ** mean significant difference from wild-type at P < 0.05 and P < 0.01, respectively.
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| 27 | (Managing Editor: Wang Caihong) |
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