Billakurthi Kumari, Hibberd Julian M
Department of Plant Sciences, University of Cambridge, Downing Street, Cambridge, UK.
Plant Methods. 2023 Jul 6;19(1):69. doi: 10.1186/s13007-023-01041-x.
It has been proposed that engineering the C photosynthetic pathway into C crops could significantly increase yield. This goal requires an increase in the chloroplast compartment of bundle sheath cells in C species. To facilitate large-scale testing of candidate regulators of chloroplast development in the rice bundle sheath, a simple and robust method to phenotype this tissue in C species is required.
We established a leaf ablation method to accelerate phenotyping of rice bundle sheath cells. The bundle sheath cells and chloroplasts were visualized using light and confocal laser microscopy. Bundle sheath cell dimensions, chloroplast area and chloroplast number per cell were measured from the images obtained by confocal laser microscopy. Bundle sheath cell dimensions of maize were also measured and compared with rice. Our data show that bundle sheath width but not length significantly differed between C rice and C maize. Comparison of paradermal versus transverse bundle sheath cell width indicated that bundle sheath cells were intact after leaf ablation. Moreover, comparisons of planar chloroplast areas and chloroplast numbers per bundle sheath cell between wild-type and transgenic rice lines expressing the maize GOLDEN-2 (ZmG2) showed that the leaf ablation method allowed differences in chloroplast parameters to be detected.
Leaf ablation is a simple approach to accessing bundle sheath cell files in C species. We show that this method is suitable for obtaining parameters associated with bundle sheath cell size, chloroplast area and chloroplast number per cell.
有人提出将C光合途径导入C作物中可显著提高产量。这一目标需要增加C植物中维管束鞘细胞的叶绿体区室。为便于对水稻维管束鞘中叶绿体发育的候选调节因子进行大规模测试，需要一种简单且可靠的方法来对C植物中的该组织进行表型分析。
我们建立了一种叶片切除方法来加速水稻维管束鞘细胞的表型分析。使用光学显微镜和共聚焦激光显微镜观察维管束鞘细胞和叶绿体。从共聚焦激光显微镜获得的图像中测量维管束鞘细胞尺寸、叶绿体面积和每个细胞的叶绿体数量。还测量了玉米的维管束鞘细胞尺寸并与水稻进行比较。我们的数据表明，C水稻和C玉米之间维管束鞘宽度存在显著差异，而长度无显著差异。比较表皮下与横向维管束鞘细胞宽度表明，叶片切除后维管束鞘细胞完整。此外，对野生型和表达玉米GOLDEN-2（ZmG2）的转基因水稻品系之间每个维管束鞘细胞的平面叶绿体面积和叶绿体数量进行比较表明，叶片切除方法能够检测到叶绿体参数的差异。
叶片切除是一种在C植物中获取维管束鞘细胞列的简单方法。我们表明该方法适用于获取与维管束鞘细胞大小、叶绿体面积和每个细胞的叶绿体数量相关的参数。
