Improved DNA barcoding method for Bemisia tabaci and related Aleyrodidae: development of universal and Bemisia tabaci biotype-specific mitochondrial cytochrome c oxidase I polymerase chain reaction primers.

Whiteflies, heteropterans in the family Aleyrodidae, are globally distributed and severe agricultural pests. The mitochondrial cytochrome c oxidase I (mtCOI) sequence has been used extensively in whitefly phylogenetic comparisons and in biotype identification of the agriculturally important Bemisia tabaci (Gennadius) whitefly. Because of the economic importance of several whitefly genera, and the invasive nature of the B and the Q biotypes of Bemisia tabaci, mtCOI sequence data are continually generated from sampled populations worldwide. Routine phylogenetic comparisons and biotype identification is done through amplification and sequencing of an approximately 800-bp mtCOI DNA fragment. Despite its routine use, published primers for amplification of this region are often inefficient for some B. tabaci biotypes and especially across whitefly species. Through new sequence generation and comparison to available whitefly mtCOI sequence data, a set of polymerase chain reaction (PCR) amplification primers (Btab-Uni primers) were identified that are more efficient at amplifying approximately 748 bp of the approximately 800-bp fragment currently used. These universal primers amplify an mtCOI fragment from numerous B. tabaci biotypes and whitefly genera by using a single amplification profile. Furthermore, mtCOI PCR primers specific for the B, Q, and New World biotypes of B. tabaci were designed that allow rapid discrimination among these biotypes. These primers produce a 478-, 405-, and 303-bp mtCOI fragment for the B, New World, and Q biotypes, respectively. By combining these primers and using rapid PCR and electrophoretic techniques, biotype determination can be made within 3 h for up to 96 samples at a time.

Spatial structure characteristic analysis of corn stover during alkali and biological co-pretreatment using XRD.

Dynamic variation in the spatial structure of corn stover during alkali and biological co-pretreatment was investigated by X-ray diffraction. The result for crystallinity and microcrystalline size of cellulose showed periodic changes during the pretreatment process. The dominant destruction periods of crystalline areas were mainly located at 3-5d and 7-17d, and prevailing destroyed amorphous areas mainly occurred at 0-2d and 5-7d. On day 7, the relative crystallinity and microcrystalline size reached 52.81% and 8.56 nm, respectively, which were the maximum and minimum values during the whole co-pretreatment. The results indicated that spatial structure change was not uniform with pretreatment time, and this was contributed to explore the vital time point of destruction during the alkali-biological pretreatment.