Identification of microRNAs involved in lipid biosynthesis and seed size in developing sea buckthorn seeds using high-throughput sequencing.

Sea buckthorn is a plant of medicinal and nutritional importance owing in part to the high levels of essential fatty acids, linoleic (up to 42%) and α-linolenic (up to 39%) acids in the seed oil. Sea buckthorn can produce seeds either via the sexual pathway or by apomixis. The seed development and maturation programs are critically dependent on miRNAs. To understand miRNA-mediated regulation of sea buckthorn seed development, eight small RNA libraries were constructed for deep sequencing from developing seeds of a low oil content line 'SJ1' and a high oil content line 'XE3'. High-throughput sequencing identified 137 known miRNA from 27 families and 264 novel miRNAs. The potential targets of the identified miRNAs were predicted based on sequence homology. Nineteen (four known and 15 novel) and 22 (six known and 16 novel) miRNAs were found to be involved in lipid biosynthesis and seed size, respectively. An integrated analysis of mRNA and miRNA transcriptome and qRT-PCR identified some key miRNAs and their targets (miR164d-ARF2, miR168b-Δ9D, novelmiRNA-108-ACC, novelmiRNA-23-GPD1, novelmiRNA-58-DGAT1, and novelmiRNA-191-DGAT2) potentially involved in seed size and lipid biosynthesis of sea buckthorn seed. These results indicate the potential importance of miRNAs in regulating lipid biosynthesis and seed size in sea buckthorn.

Changes in triacylglycerol composition during ripening of sea buckthorn (Hippophaë rhamnoides L.) seeds.

Changes in the quantitative composition of triacylglycerols (TAGs) in maturing sea buckthorn (Hippophaë rhamnoides L.) seeds were determined by lipase hydrolysis. As a whole, the rate of synthesis of separate TAG classes increased in proportion to both their unsaturation and relative content (weight percent) in total TAGs. Up to the 80th day of maturation, the formation of triunsaturated TAGs was predominant. Subsequently, at the terminal stage of seed ripening, the absolute content (in nanomoles per seed) of a major group of these TAGs containing linolenic and linoleic acyls decreased by approximately 7%, and the increase in the total TAG content was mainly due to the synthesis of TAG molecules including stearic and palmitic acyls in the rac-1,3 positions, as well as those containing oleate in the sn-2 position. At each maturation stage, the composition of the TAGs formed was controlled both by the composition of fatty acids available for TAG synthesis and by the rate of incorporation of a particular fatty acid into the sn-2 position of the TAGs.