Abstract
Kinnow mandarin (Citrus reticulata Blanco) is a popular citrus crop of northwestern India and it occupies maximum fruit area in Punjab. However, citrus juice processing industry is still suffering from delayed bitterness problem caused mainly by limonoid aglycones such as limonin. In order to study citrus limonoid metabolism, limonoid glucosyltransferase (LGT) gene, which encodes a natural debittering enzyme, was isolated from the fruit tissues of Kinnow mandarin. After confirmation and characterization, its full-length gene sequence (1533 bp) was submitted to National Centre for Biotechnology Information. Citrus reticulata limonoid glucosyltransferase (CrLGT) occupies a position on an independent branch in the largest subgroup and is phylogenetically different from those in other mandarin species like C. unshiu, showing its uniqueness in several features. The transcript expression of CrLGT, evaluated in different tissues such as young leaf, flavedo, albedo, sac covering and seed of Kinnow mandarin during early (90 days after flowering (DAF)), mid (150-210 DAF) and late (240 DAF) fruit developmental stages using semi-quantitative method, showed the highest expression in flavedo. Thus, it was concluded that the isolated LGT gene has an effect on limonoid metabolic engineering in citrus. Overexpression of this gene can reduce the delayed bitterness problem in citrus juice and enhance the accumulation of specific glucosides that have anticancer effects.
Highlights
Kinnow mandarin (Citrus reticulata Blanco) is a commercially important citrus fruit crop of northwestern India
Molecular cloning of Citrus reticulata limonoid glucosyltransferase (CrLGT) from Kinnow mandarin and its sequence comparison The quality and quantity of total RNA isolated from young leaves and fruit tissues of Kinnow mandarin were checked on 1 % denaturing agarose gel
The Citrus reticulata limonoid glucosyltransferase (CrLGT) has been reported as a key player for natural debittering and anticancer potential. This CrLGT showed several indels, absence of introns and produced a shorter translated product but with intact plant secondary product glucosyltransferase box, which is important for glycosylation
Summary
Kinnow mandarin (Citrus reticulata Blanco) is a commercially important citrus fruit crop of northwestern India. It is a hybrid between King and Willow leaf mandarin Brazil, USA, India, Mexico and Spain are the world’s leading citrus fruit producing countries and they account for two-thirds of the global production annually. India produces 10.48 million tonnes of citrus fruits and ranks fourth in production globally [1]. Kinnow mandarin occupies 61.9 % of total fruit area in Punjab and ranks first in fruit production in India [2]. Citrus industry in India has been suffering from delayed bitterness problem caused by limonoids, mainly limonin [3]. Juice producers have to discard the juice, blend it with sugar solution or remove the bitter limonoids, which otherwise degrade the quality of the juice
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