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Tigernut (Cyperus esculentus L.), which belongs to the Cyperaceae family within Poales, is a novel herbaceous oil crop producing high amounts of oil in underground tubers. Like other tuber and tuberous root crops, water accounts for approximately 85% of immature tubers, implying a crucial role of water balance for tuber development in tigernut. Plasma membrane intrinsic proteins (PIPs), which include two groups (i.e. PIP1 and PIP2) typically localized to the cell membrane, constitute a subfamily of aquaporin facilitating the passive transport of water. Based on one PIP identified in the proteomes of tigernut tubers, its coding gene CePIP1;1 was cloned using the RT-PCR technique, followed by analyzing the exon-intron structure, sequence features, evolutionary relationships, expression profiles, and protein subcellular localization. CePIP1;1 was shown to contain three introns with a coding sequence of 867 bp, putatively encoding 288 amino acids with the theoretical molecular weight of 30.76 kDa, the isoelectric point of 8.82, the instability index of 32.95, the grand average of hydropathicity of 0.384, and the aliphatic index of 95.28, implying that it is stable, basic, and hydrophobic. The protein was predicted to harbor one conserved MIP (major intrinsic protein) domain, including six transmembrane helices, two half helices, and dual NPA motifs located at the N-termini of two half helices. Sequence alignment and phylogenetic analysis revealed that CePIP1;1 clustered with Oryza sativa PIP1s and possesses an extended N-terminus but a short C-terminus relative to Spinacia oleracea PIP2;1, supporting its classification. Transient over-expression in Nicotiana benthamiana leaves supported the plasma membrane localization of CePIP1;1, which is consistent with the bioinformatics prediction. Further expression analysis showed that CePIP1;1 was constitutively expressed in all tissues examined in this study. During tuber development, CePIP1;1 exhibited a typical bell-shaped expression pattern, which is consistent with that of the moisture content. These results lay a solid foundation for further functional analysis and genetic improvement in tigernut.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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