An improved pollen number counting method using a cell counter and mesh columns
An improved pollen number counting method using a cell counter and mesh columns
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DOI：
10.1186/s13007-020-00668-4
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发表时间：
2020-09-14
期刊：
影响因子：
5.1
通讯作者：
Moriguchi, Yoshinari
中科院分区：
文献类型：
--
作者：
Kakui, Hiroyuki;Tsurisaki, Eriko;Moriguchi, Yoshinari
花粉数量的测定在进化、农业和医学研究中具有重要意义。柏科树种在世界范围内引起严重的花粉症。虽然日本雪松（柳杉）是日本最重要的林业物种，但它也是该国花粉症的最大原因。日本雪松树已被选择的生长速度和上级形态性状，然后克隆。这些无性系的花粉产量可能会有所不同，但很少有研究表明一个雄性球花（花）会产生多少花粉粒。最近报道的一种用细胞计数器计数花粉数的方法适用于松属植物和小麦，但不适用于日本雪松，因为孢子叶球不随加热（例如60 ℃，过夜）而打开。结果在这里，我们报告了一种改进的花粉计数方法，使用一个精确和快速的细胞计数器结合自制的网格柱。使用研杵轻轻压碎雄性球花。然后分别使用100-和20-μ m网孔柱去除大的和小的碎片。我们成功地检测到花粉大小和数量不同的两个克隆使用这种方法。结论改进后的方法不仅适用于日本杉花粉的计数，也适用于其它有硬鳞覆盖的柏科植物花粉的计数。此外，这种方法可以应用于更广泛的植物物种，例如小麦，因为不需要等待开花，并且可以有效地去除碎片。
Background The determination of pollen number is important in evolutionary, agricultural, and medical studies. Tree species of the Cupressaceae family cause serious pollinosis worldwide. Although Japanese cedar (Cryptomeria japonica) is the most important forestry species in Japan, it is also the biggest cause of pollinosis in the country. Japanese cedar trees have been selected for growth speed and superior morphological traits and then cloned. These clones may vary in their pollen production, but there has been little research on how many pollen grains are produced by a single male strobilus (flower). A recently reported method for counting pollen number with a cell counter was applicable toArabidopsisspecies and wheat, but was not suitable for Japanese cedar because the strobilus does not open with heating (e.g. 60 degrees C, overnight). Results Here, we report an improved pollen counting method for Japanese cedar using a precise and rapid cell counter in combination with home-made mesh columns. The male strobilus was gently crushed using a pestle. Large and small debris were then removed using 100- and 20-mu m mesh columns, respectively. We successfully detected pollen sizes and numbers that differed between two clones using this method. Conclusions This improved method is not only suitable for counting pollen from Japanese cedar, but could also be applied to other species of the Cupressaceae family with hard scale tissue covering the pollen. Moreover, this method could be applied to a broader range of plant species, such as wheat, because there is no need to wait for anthesis and debris can be removed efficiently.