Overexpression of AtATM3 in Brassica juncea confers enhanced heavy metal tolerance and accumulation

Bhuiyan, Mohammed Shafi Ullah,Min, Sung Ran,Jeong, Won Joong,Sultana, Sayeda,Choi, Kwan Sam,Lee, Youngsook,Liu, Jang R. Springer-Verlag 2011 Plant cell, tissue and organ culture Vol.107 No.1

An improved method for <i>Agrobacterium</i>-mediated genetic transformation from cotyledon explants of <i>Brassica juncea</i>

Bhuiyan, Mohammed Shafi Ullah,Min, Sung Ran,Jeong, Won Joong,Sultana, Sayeda,Choi, Kwan Sam,Lim, Yong Pyo,Song, Won Yong,Lee, Youngsook,Liu, Jang R. Japanese Society for Plant Cell and Molecular Biol 2011 Plant biotechnology Vol.28 No.1

<P>An efficient <I>Agrobacterium</I>-mediated genetic transformation method was established for <I>Brassica juncea</I> by investigating several factors responsible for successful gene transfer. Four-day-old cotyledon explants from <I>in vitro</I> grown seedlings were co-cultivated with <I>Agrobacterium</I> strain GV3101 harboring the binary vector EnPCAMBIA1302-YCF1, which contained the hygromycin phosphotransferase (<I>HPT</I>) gene as a selectable marker and the yeast cadmium factor 1 (<I>YCF1</I>) gene. Two days co-cultivation period on shoot induction medium (MS medium supplemented with 0.1 mg l<SUP>−1</SUP> α-naphthaleneacetic acid, 1.0 mg l<SUP>−1</SUP> 6-benzyladenine, and 2.0 mg l<SUP>−1</SUP> silver nitrate) containing 20 mg l<SUP>−1</SUP> acetosyringone and five days delaying exposure of explants to selective agent enhanced transformation efficiency significantly. A three-step selection strategy was developed to select hygromycin resistant shoots. Hygromycin-resistant shoots were subsequently rooted on root induction medium. Rooted plantlets were transferred to pot-soil, hardened, and grown in a greenhouse until maturity. Using the optimized transformation procedure, transformation efficiency reached at 16.2% in this study. Southern blot analysis was performed to confirm that transgenes (<I>HPT</I> and <I>YCF1</I>) were stably integrated into the plant genome. All transgenic plants showed single-copy of transgene integration in the host genome. Segregation analysis of T<SUB>1</SUB> progeny showed that the transgenes were stably integrated and transmitted to the progeny in a Mendelian fashion.</P>

Factors for high frequency plant regeneration in tissue cultures of Indian mustard (Brassica juncea L.)

Mohammed Shafi Ullah Bhuiyan,민성란,최관삼,임용표,유장렬 한국식물생명공학회 2009 식물생명공학회지 Vol.36 No.2

An efficient system for high frequency plant regeneration was established through investigating various factors such as plant growth regulator combinations, explant types and ages, and addition of AgNO3 influenced on shoot regeneration in Brassica juncea L. cv. BARI sarisha-10. Murashige and Skoog (MS) medium supplemented with 0.1 mg/L NAA (1-naphthaleneacetic acid) and 1 mg/L BA (6-benzyladenine) showed the maximum shoot regeneration frequency (56.67%) among the different combinations of NAA and BA. Explant type, explant age, and addition of AgNO3 also significantly affected shoot regeneration. Of the four type of explants (cotyledon, hypocotyl, root, and leaf explants)- cotyledon explants produced the highest shoot regeneration frequency and hypocotyls explants produced the highest number of shoots per explant, whereas root explants did not produce any shoot. The cotyledonary explants from Four-day-old seedlings showed the maximum shoot regeneration frequency and number of shoots per explant. Shoot regeneration frequency increased significantly by adding AgNO3 to the medium. Two mg/L AgNO3 appeared to be the best for shoot regeneration with the highest shoot regeneration frequency (86.67%) and number of shoots per explant (7.5 shoots). Considerable variation in shoot regeneration from cotyledonay explants was observed within the B. juncea L. genotypes. The shoot regeneration frequency ranged from 47.78% for cv. Shambol to 91.11% for cv. Rai-5. In terms of the number of shoots produced per explant, B. juncea L. cv. Daulot showed the maximum efficiency. MS medium supplemented with 0.1 mg/L NAA showed the highest frequency of rooting. The regenerated plantlets were transferred to pot soil and grown to maturity in the greenhouse. All plants were fertile and morphologically identical with the source plants.

Factors for high frequency plant regeneration in tissue cultures of Indian mustard (Brassica juncea L.)

Bhuiyan, Mohammed Shafi Ullah,Min, Sung-Ran,Choi, Kwan-Sam,Lim, Yong-Pyo,Liu, Jang-Ryol The Korean Society of Plant Biotechnology 2009 식물생명공학회지 Vol.36 No.2

An efficient system for high frequency plant regeneration was established through investigating various factors such as plant growth regulator combinations, explant types and ages, and addition of $AgNO_3$ influenced on shoot regeneration in Brassica juncea L. cv. BARI sarisha-10. Murashige and Skoog (MS) medium supplemented with 0.1 mg/L NAA (1-naphthaleneacetic acid) and 1 mg/L BA (6-benzyladenine) showed the maximum shoot regeneration frequency (56.67%) among the different combinations of NAA and BA. Explant type, explant age, and addition of $AgNO_3$ also significantly affected shoot regeneration. Of the four type of explants (cotyledon, hypocotyl, root, and leaf explants)- cotyledon explants produced the highest shoot regeneration frequency and hypocotyls explants produced the highest number of shoots per explant, whereas root explants did not produce any shoot. The cotyledonary explants from Four-day-old seedlings showed the maximum shoot regeneration frequency and number of shoots per explant. Shoot regeneration frequency increased significantly by adding $AgNO_3$ to the medium. Two mg/L $AgNO_3$ appeared to be the best for shoot regeneration with the highest shoot regeneration frequency (86.67%) and number of shoots per explant (7.5 shoots). Considerable variation in shoot regeneration from cotyledonay explants was observed within the B. juncea L. genotypes. The shoot regeneration frequency ranged from 47.78% for cv. Shambol to 91.11% for cv. Rai-5. In terms of the number of shoots produced per explant, B. juncea L. cv. Daulot showed the maximum efficiency. MS medium supplemented with 0.1 mg/L NAA showed the highest frequency of rooting. The regenerated plantlets were transferred to pot soil and grown to maturity in the greenhouse. All plants were fertile and morphologically identical with the source plants.

High frequency plant regeneration from transverse thin cell layers (tTCLs) in Indian mustard (Brassica juncea L.)

Mohammed Shafi Ullah Bhuiyan,임용표,민성란,최관삼,유장렬 한국식물생명공학회 2009 식물생명공학회지 Vol.36 No.1

An efficient and reproducible plant regeneration system was established using transverse thin cell layers (tTCLs) in five cultivars of Brassica juncea L. The effects of medium conditions, explant types (tTCLs of hypcotyl and cotyledonary petiole) on shoot regeneration were examined in this study. The maximum shoot regeneration frequency was obtained in Murashige and Skoog (MS) medium supplemented with 4 mg/L 6-benzylaminopurine (BA) and 0.2 mg/L 1-naphthaleneacetic acid (NAA). The hypocotyls derived tTCL explants had more shoot regeneration frequency (52%) than the cotyledonary petiole derived tTCL explants. Shoot induction was further improved by the addition of silver nitrate (AgNO3) in the regeneration medium. A significant genotypic effect was also observed between the five cultivars; Rai-5 displayed higher capacities to produce shoots than other cultivars. Regenerated shoots were rooted on MS basal medium without PGRs which induced 90% of roots. The plantlets established in greenhouse conditions with 99% survival, flowered normally and set seeds. The regenerated plants were fertile and identical to source plants.

Mode of Organogenesis of Epiphyllous Bud in Kalanchoe daigremontiana During In Vitro Culture System

Mohammed Shafi Ullah Bhuiyan,Kim, Tehryung,Choi, Kwan Sam 충남대학교 생물공학연구소 2005 생물공학연구지 Vol.11 No.2

Extensive segmentation of detached leaves bearing variable number of dormant buds has been done to investigate the mode of organogenesis of epiphyllous buds on the leaves in Kalanchoe daigremontiana. The study revealed that shoot always emerged prior to root emergence from each bud. The buds from the middle region of the leaves were more responsive (earlier organogenesis) and more uniformly so than those from the distal regions. The existence of such a pattern of organogenesis even in leaf cuttings and squares points to the operation of some endogenous physiological gradients along linear leaf axis, not disturbed by system miniaturization. Enlargement of photosynthetic tissue at the disposal of a single bud did not accelerate organogenesis.