In Vitro Regeneration Potential of White Lupin (Lupinus albus) from Cotyledonary Nodes.

Mehtab Muhammad Aslam,Rui Miao,Feiyun Xu,Jianping Liu,Qian Zhang,Kashif Akhtar,Huifeng Lin,Weifeng Xu,Joseph K Karanja,Tianyu Xia

doi:10.3390/plants9030318

Mehtab Muhammad Aslam, Rui Miao + Show 8 more

Open Access

https://doi.org/10.3390/plants9030318

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Abstract

The tissue culture regeneration system of Lupinus albus has always been considered as recalcitrant material due to its genotype-dependent response and low regeneration efficiency that hamper the use of genetic engineering. Establishment of repeatable plant regeneration protocol is a prerequisite tool for successful application of genetic engineering. This aim of this study was to develop standardized, efficient protocol for successful shoot induction from cotyledonary node of white lupin. In this study, 5 day old aseptically cultured seedlings were used to prepare three explants (half cotyledonary node, HCN; whole cotyledonary node, WCN; and traditional cotyledonary node, TCN), cultured on four concentrations of M519 medium (M519, ½ M519, 1/3 M519, and ¼ M519), containing four carbohydrate sources (sucrose, fructose, maltose, and glucose), and stimulated with various combinations of KT (kinetin), and NAA (naphthalene acetic acid) for direct shoot regeneration. High frequency of 80% shoot regeneration was obtained on ½ M519 medium (KT 4.0 mg L−1 + NAA 0.1 mg L−1) by using HCN as an explant. Interestingly, combinations of (KT 4.0 mg L−1 + NAA 0.1 mg L−1 + BAP 1.67 mg L−1), and (KT 2.0 mg L−1 + NAA 0.1 mg L−1) showed similar shoot regeneration frequency of 60%. Augmentation of 0.25 g L−1 activated charcoal (AC) not only reduced browning effect but also improved shoot elongation. Among the all carbohydrate sources, sucrose showed the highest regeneration frequency with HCN. Additionally, 80% rooting frequency was recorded on ½ M519 containing IAA 1.0 mg L−1 + KT 0.1 mg L−1 (indole acetic acid) after 28 days of culturing. The present study describes establishment of an efficient and successful protocol for direct plant regeneration of white lupin from different cotyledonary nodes.

Highlights

White lupin (Lupinus albus L.) is an important legume plant and produces high protein content seeds, dietary fiber, starch or gluten free, [1] and have good nutritional value [1,2]
The highest percentage of rooting was obtained on 12 M519 medium supplemented with 3% sucrose and Indole acetic acid (IAA) 1.0 mg L−1 + KT 0.1 mg L−1 (Figure 4h), while IAA alone 1.0mg L−1 showed (66%) rooting percentage
The current research was designed to evaluate the regeneration potential of Lupinus albus belonging to the family Fabaceae

Summary

Introduction

White lupin (Lupinus albus L.) is an important legume plant and produces high protein content seeds, dietary fiber, starch or gluten free, [1] and have good nutritional value [1,2]. White lupin, is capable of fixing atmospheric N2 into accessible form for plants [3]. White lupin is a cluster root (CR) forming legume and illuminated as model plant to examine morphological and biochemical adaptations of plants to phosphate deficient soils [4], and plays a key role in improving soil exploration and phosphate availability [5,6,7]. White lupin has generated heightened interest among researchers aiming to generate P-efficient genetically modified improved plants, due to the fact of its versatile application in agriculture. Most of the legume species [11,12], especially white lupin, are considered as recalcitrant material for genetic manipulation

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