Scientia Agricultura Sinica ›› 2020, Vol. 53 ›› Issue (16): 3225-3234.doi: 10.3864/j.issn.0578-1752.2020.16.003

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

Creating a New-Type Brassica napus (AnArCnCo) with High Drought-resistance Employing Hexaploid (AnAnCnCnCoCo) as a Bridge

WAN HuaFang(),WEI Shuai(),FENG YuXia,QIAN Wei()   

  1. College of Agronomy and Biotechnology, Southwest University/Academy of Agricultural Sciences, Southwest University, Chongqing 400716
  • Received:2019-10-05 Accepted:2020-02-21 Online:2020-08-16 Published:2020-08-27
  • Contact: Wei QIAN E-mail:wanhua05@163.com;499623219@qq.com;qianwei666666@hotmail.com

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Abstract:

【Objective】As one parent of Brassica napus, B. rapa has a wide range of variation, as well as excellent performance against drought or poor soil. Employing hexaploid as a bridge, the drought resistance of B. rapa is transferred to develop a new-type B. napus with excellent drought tolerance.【Method】The hexaploid (AnAnCnCnCoCo) was created via interspecific cross between B. napus (AnAnCnCn) and B. oleracea (CoCo) via embryo rescue and chrosome doubling. Next, a new-type B. napus（AnArCnCo）was developed by crossing the hexaploid with B. rapa (ArAr). Under the treatment with PEG-6000 solution to imitate drought stress at germination stage, the indices related to drought resistance were measured in the experimental materials, including natural B. napus, new-type B. napus and B.rapa. All the datum were analyzed with subordinate function method.【Result】 Using 11 hexaploid materials and 68 B. rapa materials as parents, 124 new-type B. napus lines were created, which showed intermediate morphology between hexaploid and B. rapa. The hexaploid used in the experiment all had 56 chrosomes and the new-type B. napus had 38 chromsomes. The pollen fertility in the hexaploid and the new-type B. napus was approximately 90% and 80%, respectively. Based on the results of principal components analysis and SSR analysis, 7 hexaploid, 59 new-type B. napus, 10 B. rapa and 20 natural B. napus were clustered into three groups, namely B.rapa, hexaploid and B. napus. The new-type B. napus and natural B. napus were in the same group, but they clearly separated each other. Based on the drought resistance-related indices, 200 g·L-1 and 250 g·L-1 PEG-6000 solution were used to imitate drought stress to B. rapa and new-type B. napus, respectively. Nine new-type B. napus were tested, but only three were more resistant than the control, Zhongshuang 11. Additionally, the resistance were positively associated to the corresponding parent B. rapa, which indicated that the three new-type B. napus have been transfered excellent drought resistance from B. rapa.【Conclusion】 Using B. rapa as a drought resistance donor and the hexaploid as a bridge, new-type B. napus lines with excellent drought tolerance were developed.

Key words: Brassica napus, Brassica rapa, drought tolerance, PEG-6000, principal components analysis

Fig. 1

Identification of new-type B. napus Ⅰ: Seedling phenotype of hexaploid (AnAnCnCnCoCo); Ⅱ: Seedling phenotype of the new-type B. napus; Ⅲ: Seedling phenotype of B. rapa; Ⅳ: Chrosomes of hexaploid; Ⅴ: Chrosomes of new-type B. napus; Ⅵ: Pollen stainability of hexaploid; Ⅶ: Pollen stainability of new-type B. napus; Ⅷ: Principal component analysis result, “round” refers to B. rapa; “red square” refers to a hexaploid; “green diamond” refers to new-type B. napus; “blue triangle” refers to natural B. napus"

Fig. 2

Root length of rapeseed treated by PEG-6000 solution with different concentration Ⅰ: Average root length of new-type B. napus treated by PEG-6000 solution with different concentration;Ⅱ: Variation coefficient of relative root length of the new-type B. napus treated by PEG-6000 solution with different concentration;Ⅲ: Average root length of B. rapa treated by PEG-6000 solution with different concentration; Ⅳ: Variation coefficient of relative root length of B. rapa treated by PEG-6000 solution with different concentration. Different letters indicate significant difference at P<0.01"

Table 1

Primitive matrix of principal component analysis on drought-resistance indices of rapeseed at germination stage"

编号
Code		 类型
Type		 萌发抗旱指数
DRI		 相对萌发率
RGR		 相对萌发势
RSP		 相对根长
RRL		 相对胚轴长
RSL
9X006 新型甘蓝型油菜New-type B. napus 0.262 0.067 0.069 0.209 0.106
9X002 新型甘蓝型油菜New-type B. napus 0.543 0.067 0.000 0.690 0.133
9X025 新型甘蓝型油菜New-type B. napus 0.798 0.300 0.296 0.718 0.135
9X016 新型甘蓝型油菜New-type B. napus 0.392 0.600 0.615 0.319 0.173
9X018 新型甘蓝型油菜New-type B. napus 0.824 0.900 0.827 0.692 0.249
9X111 新型甘蓝型油菜New-type B. napus 0.483 0.633 0.393 0.748 0.162
9X028 新型甘蓝型油菜New-type B. napus 0.591 0.933 0.933 0.156 0.109
9X015 新型甘蓝型油菜New-type B. napus 0.637 0.600 0.533 0.228 0.151
9X026 新型甘蓝型油菜New-type B. napus 0.380 0.700 0.633 0.122 0.109
中双11 ZS11 甘蓝型油菜B. napus 0.800 0.867 0.700 0.185 0.117
8M664 白菜型油菜B. rapa 0.800 0.830 0.860 0.690 0.210
8M681 白菜型油菜B. rapa 0.670 0. 830 0.800 0.920 0.410
8M625 白菜型油菜B. rapa 0.530 0.570 0.530 0.310 0.260
8M623 白菜型油菜B. rapa 0.520 0.900 0.830 0.610 0.500
8M684 白菜型油菜B. rapa 0.290 0.470 0.410 0.560 0.130
8M624 白菜型油菜B. rapa 0.370 0.700 0.500 0.510 0.210
8M693 白菜型油菜B. rapa 0.450 0.730 0.730 0.690 0.300
8M191 白菜型油菜B. rapa 0.300 0.600 0.480 0.500 0.810
8M655 白菜型油菜B. rapa 0.420 0.730 0.660 0.690 0.250

Table 2

Principal component analysis on drought-resistance-related indicators on rapeseed"

主成分
Principal component		 类别
Category		 特征值
Eigen value		 贡献率Contribution ratio (%) 累计贡献率Accumulated contribution ratio (%) 抗旱指标特征向量Eigen vector of measured indicators
萌发抗旱指数DRI 相对萌发率RGR 相对萌发势RSP 相对根长RRL 相对胚轴长RSL
PC1 新型甘蓝型油菜
New-type B. napus		 2.431 48.620 48.620 0.416 0.597 0.589 -0.024 0.350
PC2 新型甘蓝型油菜
New-type B. napus		 1.774 35.470 84.090 0.332 -0.224 -0.274 0.720 0.497
PC3 新型甘蓝型油菜
New-type B. napus		 0.597 11.930 96.020 -0.784 0.083 0.110 0.011 0.605
PC1 白菜型油菜
B. rapa		 3.580 71.610 71.600 0.459 0.512 0.520 0.468 0.194
PC2 白菜型油菜
B. rapa		 0.950 19.010 90.600 -0.313 0.08 -0.073 -0.038 0.946
PC3 白菜型油菜
B. rapa		 0.320 6.430 97.000 0.599 -0.001 0.103 -0.775 0.175

Table 3

Membership function value of drought-resistance-related indicators of rapeseed under simulated drought stress"

编号
Code		 类型
Type		 萌发抗旱指数
DRI		 相对萌发率
RGR		 相对萌发势RSP 相对根长
RRL		 相对胚轴长
RSL		 均值
Means
9X006 新型甘蓝型油菜New-type B. napus 1.00 0.00 0.01 0.73 0.20 0.388
9X002 新型甘蓝型油菜New-type B. napus 0.78 0.09 0.00 1.00 0.19 0.412
9X025 新型甘蓝型油菜New-type B. napus 1.00 0.25 0.24 0.88 0.00 0.474
9X026 新型甘蓝型油菜New-type B. napus 0.46 1.00 0.89 0.02 0.00 0.474
9X028 新型甘蓝型油菜New-type B. napus 0.58 1.00 1.00 0.05 0.00 0.526
9X111 新型甘蓝型油菜New-type B. napus 0.55 0.80 0.39 1.00 0.00 0.548
中双11 ZS11 甘蓝型油菜B. napus 0.91 1.00 0.78 0.09 0.00 0.556
9X016 新型甘蓝型油菜New-type B. napus 0.50 0.96 1.00 0.33 0.00 0.558
9X015 新型甘蓝型油菜New-type B. napus 1.00 0.92 0.78 0.15 0.00 0.570
9X018 新型甘蓝型油菜New-type B. napus 0.88 1.00 0.89 0.68 0.00 0.690
8M623 白菜型油菜B. rapa 0.07 1.00 0.83 0.28 0.00 0.436
8M191 白菜型油菜B. rapa 0.00 0.61 0.35 0.40 1.00 0.472
8M624 白菜型油菜B. rapa 0.31 1.00 0.59 0.62 0.00 0.504
8M684 白菜型油菜B. rapa 0.37 0.78 0.66 1.00 0.00 0.562
8M625 白菜型油菜B. rapa 0.88 1.00 0.89 0.17 0.00 0.588
8M681 白菜型油菜B. rapa 0.51 0.83 0.76 1.00 0.00 0.620
8M655 白菜型油菜B. rapa 0.35 1.00 0.84 0.91 0.00 0.620
8M693 白菜型油菜B. rapa 0.36 1.00 1.00 0.90 0.00 0.652
8M664 白菜型油菜B. rapa 0.90 0.96 1.00 0.73 0.00 0.721

Table 4

Correlation analysis of simulated drought membership function values"

编号Code 9X006 9X002 9X025 9X026 9X028 9X111 中双11 ZS 11 9X016 9X015 9X018
8M623 -0.686 -0.583 -0.336 0.874* 0.833* 0.354 0.610 0.909* 0.522 0.666
8M191 -0.631 -0.497 -0.821* -0.274 -0.362 -0.475 -0.563 -0.398 -0.665 -0.734
8M624 -0.309 -0.129 0.081 0.692 0.651 0.764 0.560 0.791 0.503 0.789
8M684 0.023 0.289 0.398 0.311 0.300 0.912* 0.216 0.539 0.201 0.695
8M625 -0.151 -0.317 0.122 0.923** 0.948** 0.265 0.992** 0.891* 0.986** 0.872*
8M681 0.064 0.28 0.455 0.400 0.402 0.906* 0.341 0.622 0.347* 0.793
8M655 -0.232 -0.005 0.177 0.569 0.547 0.805 0.409 0.748 0.372 0.775
8M693 -0.291 -0.089 0.122 0.624 0.611 0.721 0.440 0.805 0.404 0.784
8M664 0.067 0.052 0.441 0.742 0.778 0.651 0.800 0.864* 0.818* 0.991**
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