Investigation of the Efficiency of Direct and Indirect Regeneration in Evening Primrose (Oenothera biennis)

Reza Faramarzi Hafez,Bahram Heidari,Zeynab Shahabzadeh,Bahram. Heidari 한국작물학회 2013 Journal of crop science and biotechnology Vol.16 No.4

As a medicinal plant, the importance of evening primrose (Oenothera biennis L.) is due to its unsaturated fatty acids in the seedsand roots, and also oenotherine and comfarol in the leaves. Low germination and difficulties in seed production are the main problemsencountered with growing this plant in the field. As an alternative approach, an in vitro experiment was set up for the evaluationof evening primrose production via direct and indirect regeneration of the cultivars NC-1 and VNK. For callogenesis and directregeneration, the explants from the apical bud and petiole were cultured on MS medium supplemented with 0.25, 0.75, and 1.25 mgL-1 of both BAP and Kinetin (KIN). Indirect regeneration was performed by placing apical buds, petioles, and leaf explants on MSmedium supplemented with 0.5 and 1 mg L-1 2,4-D and 0.5, 1, and 1.25 mg L-1 of both BAP and KIN. The highest shoot inductionfrom direct regeneration was obtained with apical bud explants of VNK treated with 0.75 mg L-1 BAP. The highest callus weight(3.17 g) obtained from indirect regeneration was with petiole explants treated with 1 mg L-1 2, 4-D and 1 mg L-1 BAP in VNK cultivars. The highest number of torpedo embryogenic clusters (23.8) was obtained from the VNK petiole explants treated with 0.5 mg L-12, 4-D and 1.25 mg L-1 BAP. BAP had higher positive effects on in vitro production of evening primrose than KIN in both direct andindirect regeneration. In general, results indicated that VNK was more potent for regeneration than NC-1 and concentrations of 0.75mg L-1 BAP for direct and 0.5 mg L-1 2, 4-D and 1.25 mg L-1of BAP for indirect regeneration had a higher efficiency for increasing invitro production of evening primrose

Analysis of genetic diversity in a collection of Plantago species: application of ISSR markers

Bagheri Motahareh,Heidari Bahram,Dadkhodaie Ali,Heidari Zahra,Daneshnia Negar,Richards Christopher M. 한국작물학회 2022 Journal of crop science and biotechnology Vol.25 No.1

The analysis of genetic diversity in medicinal plant species can greatly facilitate germplasm conservation and selection for use in breeding schemes. The aim of the present study was to assess the genetic diversity and diferentiation of eight Plantago species using inter simple sequence repeat (ISSR) markers. Thirty-one Plantago accessions belonging to eight species were collected from Iran environments. Polymorphisms in the DNA of Plantago accessions were analyzed using 25 ISSR primers. The data for 286 polymorphic bands indicated that the ISSR primers each amplifed 5–21 polymorphic amplicons that were 100–3000 bp in size. The mean polymorphism was 83.83% and fve primers showed higher polymorphism. The polymorphism information content (PIC) ranged from 0.1103 to 0.3829 with a mean of 0.2727. Accessions in P. amplexicaulis and P. pysillum represented the highest Nei’s and Shannon’s genetic diversity, while the lowest was obtained for P. lagopus. The phenetic network generated by the Neighbor-Net algorithm showed moderate confict and splitting of the eight species. Principal coordinate analysis (PCoA) results showed the separation of accessions of the eight species. The results suggest that ISSR markers can diferentiate Plantago species and that information on genetic diversity may assist in pre-breeding evaluations and Plantago germplasm conservation.

Investigation forMaize Inbred Lines Resistance to Maize Rough Dwarf Virus(MRDV)

Afshar. Estakhr,Bahram. Heidari,Ali Dadkhodaie,Mahmood Masoumi,Hassan Pakniyat,Zahra Ahmadi 한국작물학회 2016 Journal of crop science and biotechnology Vol.19 No.1

Maize rough dwarf virus (MRDV) is the most important disease affecting maize growth in Iran. In the present study, the putative resistance capacity of 35 maize inbred lines against the causal agents of MRDVwas studied under natural field infection in 2010 and 2011. The viral transmitter was a plant hopper known as Laodelphax steriatellus. Early sowing provided suitable temperatures for vector propagation and transmission and therefore natural infection under field conditions. Sowing single rows of SC704 as vector spreader between every five rows of inbred lines caused adequate vector propagation and viral transmission. This shows that viral infection was sufficient and possible resistance was linked to viral resistance, not failure, in vector transmission. In principal component biplot, plant height vector had wide angles with the vectors for disease severity index and MRDV incidence that showed the negative association of plant height with MRDV disease. The percentage of natural infection to MRDV in susceptible control(SC704) was about 64%. The genotypes A679, K3547/3, K3545/6, K3653/2, and B84 were more susceptible to MRDV. Only the line K3640/3 had low disease incidence and severity and it can be considered as resistant for possible use in genetic studies. Results showed that an early sowing of maize in temperate regions increases the chance of transmitting viruses via their vector.

Regenerating Salt Tolerant Saffron (Crocus sativus) Using Tissue Culture with Increased Pharmaceutical Ingredients

Zeynab Shahabzadeh,Bahram. Heidari,Ali Dadkhodaie 한국작물학회 2013 Journal of crop science and biotechnology Vol.16 No.3

Due to its vegetative reproduction, saffron has a narrow genetic base and induced in vitro variations provide opportunities for expanding new cultivars. The objectives of this study were to evaluate sodium azide-induced variations in saffron’s corm culture in order to increase salt tolerance and pharmaceutical ingredients. Corm explants from the well-known ecotypes, Estahban and Kashmar, were subjected to various concentrations of sodium azide (NaN3) (0.09, 0.12, and 0.22 mg L-1) and NaCl (1.5, 2.5, and 4.0dS equivalent to 0.07, 0.12, and 0.20 g NaCl in 100 mL water) in Murashige and Skoog medium supplemented with 1 mg L-1 2-4-D,1 mg L-1 BAP, and 30 g L-1 sucrose and in a second pot culture experiment. The active pharmaceutical ingredients (crocin, picrocrocin,and safranal) were measured by high-performance liquid chromatography (HPLC). Variations in sodium azide-treated plants were more broadened for callus fresh weight (0.57 - 7.57 g), embryo weight (1.24 - 10.29 g), and regenerated seedlings (3.0 - 21.25)compared with those (0.12 - 3.77 g, 0.56 - 4.56 g, and 0.25 - 11.50, respectively) that were not treated with sodium azide. Under 0.20% salt, flowering failed in some of plants developed from sodium azide-untreated corms. HPLC analysis indicated wider ranges for crocin (11.92 - 18.03 mg g-1), picrocrocin (8.99 - 14.76 mg g-1), and safranal (2.13 - 7.36 mg g-1) in sodium azide-treated plants compared to the ranges (0.0 - 16.1, 0.0 - 12.5, and 0.0 - 6.66 mg g-1, respectively) in untreated plants. From a breeding perspective,induced variations found in this study would be useful to improve saffron's quality and salt tolerance

Regenerating Salt Tolerant Saffron (Crocus sativus) Using Tissue Culture with Increased Pharmaceutical Ingredients

Shahabzadeh, Zeynab,Heidari, Bahram,Dadkhodaie, Ali 한국작물학회 2013 Journal of crop science and biotechnology Vol.16 No.3

Due to its vegetative reproduction, saffron has a narrow genetic base and induced in vitro variations provide opportunities for expanding new cultivars. The objectives of this study were to evaluate sodium azide-induced variations in saffron's corm culture in order to increase salt tolerance and pharmaceutical ingredients. Corm explants from the well-known ecotypes, Estahban and Kashmar, were subjected to various concentrations of sodium azide ($NaN_3$) (0.09, 0.12, and 0.22 mg $L^{-1}$) and NaCl (1.5, 2.5, and 4.0 dS equivalent to 0.07, 0.12, and 0.20 g NaCl in 100 mL water) in Murashige and Skoog medium supplemented with 1 mg $L^{-1}$ 2-4-D, 1 mg $L^{-1}$ BAP, and 30 g $L^{-1}$ sucrose and in a second pot culture experiment. The active pharmaceutical ingredients (crocin, picrocrocin, and safranal) were measured by high-performance liquid chromatography (HPLC). Variations in sodium azide-treated plants were more broadened for callus fresh weight (0.57 - 7.57 g), embryo weight (1.24 - 10.29 g), and regenerated seedlings (3.0 - 21.25) compared with those (0.12 - 3.77 g, 0.56 - 4.56 g, and 0.25 - 11.50, respectively) that were not treated with sodium azide. Under 0.20% salt, flowering failed in some of plants developed from sodium azide-untreated corms. HPLC analysis indicated wider ranges for crocin (11.92 - 18.03 mg $g^{-1}$), picrocrocin (8.99 - 14.76 mg $g^{-1}$), and safranal (2.13 - 7.36 mg $g^{-1}$) in sodium azide-treated plants compared to the ranges (0.0 - 16.1, 0.0 - 12.5, and 0.0 - 6.66 mg $g^{-1}$, respectively) in untreated plants. From a breeding perspective, induced variations found in this study would be useful to improve saffron's quality and salt tolerance.

Induction of Transgenic Hairy Roots in Trigonella foenumgraceum Co-cultivated with Agrobacterium rhizogenes Harboring a GFP Gene

Zeynab Shahabzadeh,Bahram. Heidari,Reza Faramarzi Hafez 한국작물학회 2013 Journal of crop science and biotechnology Vol.16 No.4

An important feature of A. rhizogenes-induced hairy roots is their unique ability for investigation of gene function and productionof secondary metabolites such as diosgenin in fenugreek. In order to evaluate the transformation frequency and the efficiency oftransgenic hairy root induction, leaf and stem explants from two fenugreek ecotypes, Karaj and Bushehr, were infected with threeconcentrations of OD600= 0.8, 1.2, and 1.6 of A. rhizogenes strain K599 harboring a GFP gene. Regardless of ecotype, the ability ofstem explants for the induction of hairy roots (8.09) and the transformation frequency (81.3%) was higher compared with leafexplants with the values of 5.97 and 71.88%, respectively. The number of transgenic GFP-positive hairy roots ranged from 4.2 to13.5 in the Karaj ecotype and 3.8 to 9.9 in Bushehr. The highest transgenic hairy root (8.76), the transformation frequency (79.76%),and the growth rate of transgenic roots (0.77 d-1) were obtained from infection with K599 at OD600= 1.2, while the lowest belonged tothe bacterial concentration of OD600=1.6. Although the ecotype Bushehr had lower total roots (7.53) and transgenic hairy roots (6.08),it showed higher transformation frequency (79.56%) than Karaj (73.63%). Therefore, the results indicate the importance of genotype,type of explant and bacterial concentration in breeding for induction of transgenic hairy roots and consequently, production of secondarymetabolites in fenugreek.

Induction of Transgenic Hairy Roots in Trigonella foenum-graceum Co-cultivated with Agrobacterium rhizogenes Harboring a GFP Gene

Shahabzadeh, Zeynab,Heidari, Bahram,Hafez, Reza Faramarzi 한국작물학회 2013 Journal of crop science and biotechnology Vol.16 No.4

An important feature of A. rhizogenes-induced hairy roots is their unique ability for investigation of gene function and production of secondary metabolites such as diosgenin in fenugreek. In order to evaluate the transformation frequency and the efficiency of transgenic hairy root induction, leaf and stem explants from two fenugreek ecotypes, Karaj and Bushehr, were infected with three concentrations of $OD_{600}=0.8$, 1.2, and 1.6 of A. rhizogenes strain K599 harboring a GFP gene. Regardless of ecotype, the ability of stem explants for the induction of hairy roots (8.09) and the transformation frequency (81.3%) was higher compared with leaf explants with the values of 5.97 and 71.88%, respectively. The number of transgenic GFP-positive hairy roots ranged from 4.2 to 13.5 in the Karaj ecotype and 3.8 to 9.9 in Bushehr. The highest transgenic hairy root (8.76), the transformation frequency (79.76%), and the growth rate of transgenic roots ($0.77d^{-1}$) were obtained from infection with K599 at $OD_{600}=1.2$, while the lowest belonged to the bacterial concentration of $OD_{600}=1.6$. Although the ecotype Bushehr had lower total roots (7.53) and transgenic hairy roots (6.08), it showed higher transformation frequency (79.56%) than Karaj (73.63%). Therefore, the results indicate the importance of genotype, type of explant and bacterial concentration in breeding for induction of transgenic hairy roots and consequently, production of secondary metabolites in fenugreek.

Association of DREB Genes and Microsatellite Markers Linked to NAX2 with Salt Tolerance in CIMMYT-derived Triticale, Wheat and Rye Genotypes

Roghiyeh Feuj,Bahram Heidari,Ali Dadkhodaie 한국작물학회 2018 Journal of crop science and biotechnology Vol.21 No.4

Twenty-eight genotypes including triticale, wheat and rye were grown in a greenhouse under control, 14 and 21 dS/m salinity (NaCl and CaCl2, 1:1 ratio) levels to assess variations in agronomic traits, and to assess association of dehydration responsive elements (DREB genes) and five microsatellite (SSR) markers with salt tolerance. The results of grain yield variation and principal component (PC) analysis for statistical indices revealed that the first PC under 14 and 21 dS/m salinity levels was associated with salt tolerance. Five microsatellite (Xgwm291, Xgwm312, Xgwm410, Xgwm2181 and Xgwm126) and two DREB (DREB1 and DREB2) markers were used for detection of polymorphism. In total, 35 alleles ranging from 130 to 850 bp in size were identified of which, 25 alleles were found for SSR markers and 10 belonged to DREB genes. The number of alleles ranged from 3 to 9 with an average of 5 per primer. The polymorphic information content (PIC) value ranged from 0.51 (Xgwm291) to 0.77 (DREB1) with an average of 0.68. The Xgwm291 primer amplified 3 bands (130, 160 and 185 bp) in triticale and wheat but neither were detected in rye. Remarkably, the 130 bp band was amplified in TRT826 which was categorized as salt tolerant on the basis of K+/Na+ ratio and grain yield variations. Accordingly, this band could be associated with salt tolerance and its isolation and sequencing could clarify its characteristics for possible use in marker assisted breeding (MAB) for salt tolerance in triticale and wheat.

Combining Ability and Gene Action for Maturity and Agronomic Traits in Different Heterotic Groups of Maize Inbred Lines and Their Diallel Crosses

Estakhr, Afshar,Heidari, Bahram 한국작물학회 2012 Journal of crop science and biotechnology Vol.15 No.3

Combining ability information is necessary for selection of suitable advanced lines for hybridization and identification of promising hybrids for development of improved varieties. A number of 14 maize (Zea mays L.) inbred lines and 91 related crosses were evaluated over two years, 2008 and 2009, in a temperate-zone of Iran. The objectives of the study were to identify the best general and specific combiners, heterosis and type of gene actions responsible for agronomic traits. Except for grain yield and growing degree day to milky, significant general (GCA) and specific (SCA) combining ability were observed for all traits. The Baker ratio for plant height (0.15), ear height (0.26), growing degree day to milky stage (0.04), and grain yield (0.002) showed the predominance of non-additive gene effects in the expression of these traits. The heterosis observed for grain yield, grain number, pollination period, ear and plant height was considerably higher than that observed for other traits. The correlations (r) of $F_1$ means and SCA effects were positive and significantly higher than that of r ($F_1$, mid-parents) and r ($F_1$, heterosis) for all the traits except cob percent, growing degree days to silking, and physiological maturity. MO17, K3547/5, and K3615/2 had negative GCA effects for growing degree day to milky stage and maturity. Among hybrids, MO17 ${\times}$ K3653/2, B73 ${\times}$ K3651/2, and K3545/6 ${\times}$ K3493/1 with positive SCAs for pollination period and grain yield had also negative SCA effects for degree day to silking and milky stages. Therefore, the use of these inbred lines and hybrids increases the response to selection for increasing grain yield and early maturity in maize.

Combining Ability and Gene Action for Maturity and Agronomic Traits in Different Heterotic Groups of Maize Inbred Lines and Their Diallel Crosses

Afshar. Estakhr,Bahram. Heidari 한국작물학회 2012 Journal of crop science and biotechnology Vol.15 No.3

Combining ability information is necessary for selection of suitable advanced lines for hybridization and identification of promising hybrids for development of improved varieties. A number of 14 maize (Zea mays L.) inbred lines and 91 related crosses were evaluated over two years, 2008 and 2009, in a temperate-zone of Iran. The objectives of the study were to identify the best general and specific combiners, heterosis and type of gene actions responsible for agronomic traits. Except for grain yield and growing degree day to milky, significant general (GCA) and specific (SCA) combining ability were observed for all traits. The Baker ratio for plant height (0.15), ear height (0.26), growing degree day to milky stage (0.04), and grain yield (0.002) showed the predominance of non-additive gene effects in the expression of these traits. The heterosis observed for grain yield, grain number, pollination period,ear and plant height was considerably higher than that observed for other traits. The correlations (r) of F1 means and SCA effects were positive and significantly higher than that of r (F1, mid-parents) and r (F1, heterosis) for all the traits except cob percent, growing degree days to silking, and physiological maturity. MO17, K3547/5, and K3615/2 had negative GCA effects for growing degree day to milky stage and maturity. Among hybrids, MO17 × K3653/2, B73 × K3651/2, and K3545/6 × K3493/1 with positive SCAs for pollination period and grain yield had also negative SCA effects for degree day to silking and milky stages. Therefore, the use of these inbred lines and hybrids increases the response to selection for increasing grain yield and early maturity in maize.