Direct regeneration and efficient in vitro root development studies in lentil (Lens culinaris Medik)

Tasawar Sultana, Nadia Majeed, Fariha Khan, Ata Ur Rehman, S. M Saqlan Naqvi

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Abstract

Lentil is a self-pollinating annual crop with increasing demand all over the world due to its high protein content and easy digestibility. However, like many other crops lentil too needs improvement for which conventional as well as biotechnological tools are to be employed. This study was aimed at development of tissue culture protocol especially targeting improved root development to ensure their establishment in soil in order to use their potential towards genetic manipulation. Two Pakistani lentil cultivars, Masoor-2002 and Manshera-89 were used to obtain cotyledonary nodes, epicotyl and hypocotyl explants. The explants were cultured on shoot regeneration medium containing different concentration of kinetin, BAP and tyrosine with the addition of GA3, with or without charcoal for shoot development. Masoor-2002, showed the highest frequency of shoot development on MS medium containing 5.5 mg/L tyrosine, 0.25 mg/L kinetin, 1.0 mg/L BAP, 0.1 mg/L GA3, using cotyledonary node as explant. The addition of 2 g/L of charcoal in shoot medium resulted in healthier plants, but the number of shoots were reduced. Regarding the effect of age of explants on regeneration frequency, cotyledonary nodes of age 4-6 days had higher regeneration potential. Well-developed shoots were shifted to rooting medium containing different concentration of auxin with or without charcoal. Healthier and more roots were observed on medium containing 4 mg/L IAA with addition of 2 g/L charcoal. Plants were better established (70% survival) in a soil mix containing perlite, vermiculite and peat moss in 1:1:1 ratio.
Original languageEnglish
Pages (from-to)1999-2004
Number of pages6
JournalPakistan Journal of Botany
Volume48
Issue number5
Publication statusPublished - Oct 2016

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Lens culinaris
lentils
charcoal
shoots
explants
kinetin
tyrosine
epicotyls
perlite
Sphagnum
vermiculite
crops
genetic engineering
indole acetic acid
hypocotyls
tissue culture
rooting
auxins
soil
pollination

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Sultana, Tasawar ; Majeed, Nadia ; Khan, Fariha ; Rehman, Ata Ur ; Naqvi, S. M Saqlan. / Direct regeneration and efficient in vitro root development studies in lentil (Lens culinaris Medik). In: Pakistan Journal of Botany. 2016 ; Vol. 48, No. 5. pp. 1999-2004.
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abstract = "Lentil is a self-pollinating annual crop with increasing demand all over the world due to its high protein content and easy digestibility. However, like many other crops lentil too needs improvement for which conventional as well as biotechnological tools are to be employed. This study was aimed at development of tissue culture protocol especially targeting improved root development to ensure their establishment in soil in order to use their potential towards genetic manipulation. Two Pakistani lentil cultivars, Masoor-2002 and Manshera-89 were used to obtain cotyledonary nodes, epicotyl and hypocotyl explants. The explants were cultured on shoot regeneration medium containing different concentration of kinetin, BAP and tyrosine with the addition of GA3, with or without charcoal for shoot development. Masoor-2002, showed the highest frequency of shoot development on MS medium containing 5.5 mg/L tyrosine, 0.25 mg/L kinetin, 1.0 mg/L BAP, 0.1 mg/L GA3, using cotyledonary node as explant. The addition of 2 g/L of charcoal in shoot medium resulted in healthier plants, but the number of shoots were reduced. Regarding the effect of age of explants on regeneration frequency, cotyledonary nodes of age 4-6 days had higher regeneration potential. Well-developed shoots were shifted to rooting medium containing different concentration of auxin with or without charcoal. Healthier and more roots were observed on medium containing 4 mg/L IAA with addition of 2 g/L charcoal. Plants were better established (70{\%} survival) in a soil mix containing perlite, vermiculite and peat moss in 1:1:1 ratio.",
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Direct regeneration and efficient in vitro root development studies in lentil (Lens culinaris Medik). / Sultana, Tasawar; Majeed, Nadia; Khan, Fariha; Rehman, Ata Ur; Naqvi, S. M Saqlan.

In: Pakistan Journal of Botany, Vol. 48, No. 5, 10.2016, p. 1999-2004.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Direct regeneration and efficient in vitro root development studies in lentil (Lens culinaris Medik)

AU - Sultana, Tasawar

AU - Majeed, Nadia

AU - Khan, Fariha

AU - Rehman, Ata Ur

AU - Naqvi, S. M Saqlan

PY - 2016/10

Y1 - 2016/10

N2 - Lentil is a self-pollinating annual crop with increasing demand all over the world due to its high protein content and easy digestibility. However, like many other crops lentil too needs improvement for which conventional as well as biotechnological tools are to be employed. This study was aimed at development of tissue culture protocol especially targeting improved root development to ensure their establishment in soil in order to use their potential towards genetic manipulation. Two Pakistani lentil cultivars, Masoor-2002 and Manshera-89 were used to obtain cotyledonary nodes, epicotyl and hypocotyl explants. The explants were cultured on shoot regeneration medium containing different concentration of kinetin, BAP and tyrosine with the addition of GA3, with or without charcoal for shoot development. Masoor-2002, showed the highest frequency of shoot development on MS medium containing 5.5 mg/L tyrosine, 0.25 mg/L kinetin, 1.0 mg/L BAP, 0.1 mg/L GA3, using cotyledonary node as explant. The addition of 2 g/L of charcoal in shoot medium resulted in healthier plants, but the number of shoots were reduced. Regarding the effect of age of explants on regeneration frequency, cotyledonary nodes of age 4-6 days had higher regeneration potential. Well-developed shoots were shifted to rooting medium containing different concentration of auxin with or without charcoal. Healthier and more roots were observed on medium containing 4 mg/L IAA with addition of 2 g/L charcoal. Plants were better established (70% survival) in a soil mix containing perlite, vermiculite and peat moss in 1:1:1 ratio.

AB - Lentil is a self-pollinating annual crop with increasing demand all over the world due to its high protein content and easy digestibility. However, like many other crops lentil too needs improvement for which conventional as well as biotechnological tools are to be employed. This study was aimed at development of tissue culture protocol especially targeting improved root development to ensure their establishment in soil in order to use their potential towards genetic manipulation. Two Pakistani lentil cultivars, Masoor-2002 and Manshera-89 were used to obtain cotyledonary nodes, epicotyl and hypocotyl explants. The explants were cultured on shoot regeneration medium containing different concentration of kinetin, BAP and tyrosine with the addition of GA3, with or without charcoal for shoot development. Masoor-2002, showed the highest frequency of shoot development on MS medium containing 5.5 mg/L tyrosine, 0.25 mg/L kinetin, 1.0 mg/L BAP, 0.1 mg/L GA3, using cotyledonary node as explant. The addition of 2 g/L of charcoal in shoot medium resulted in healthier plants, but the number of shoots were reduced. Regarding the effect of age of explants on regeneration frequency, cotyledonary nodes of age 4-6 days had higher regeneration potential. Well-developed shoots were shifted to rooting medium containing different concentration of auxin with or without charcoal. Healthier and more roots were observed on medium containing 4 mg/L IAA with addition of 2 g/L charcoal. Plants were better established (70% survival) in a soil mix containing perlite, vermiculite and peat moss in 1:1:1 ratio.

KW - Auxin

KW - Charcoal

KW - Cytokinin

KW - Explant

KW - Lentil

KW - Regeneration

KW - Rooting

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M3 - Article

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VL - 48

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JO - Pakistan Journal of Botany

JF - Pakistan Journal of Botany

SN - 0556-3321

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