Sex determination in the early developmental stages of dioecious crops is economically-beneficial... more Sex determination in the early developmental stages of dioecious crops is economically-beneficial. During this study, a human homology of SRY gene was successfully identified in dioecious crops. SRY gene sequences of date palm and jojoba were submitted to GenBank under the accession numbers KC577225 and MK991776, respectively. This is the first report regarding the novel sex-determination methodology of four dioecious plants (jojoba, date palm, papaya, and pistachios). SRY sex gene was found in all the tested dioecious plant and human samples. This novel approach is simple and of significant importance for breeders. It facilitates the unambiguous selection of jojoba and date palm female plants at an early age and reduces the plantation cost of cultivating non-productive male plants. This is a rapid sexdetermination technique for dioecious plants and mammals at an early stage. This technique specifically targets the SRY sequence that has been comprehensively investigated in humans. The kit development for the SRY-based sex determination of various crops is in progress.
BackgroundJojoba (Simmondsia Chinensis L.) is a dioecious perennial evergreen shrub native to the... more BackgroundJojoba (Simmondsia Chinensis L.) is a dioecious perennial evergreen shrub native to the south-western desert of North America and has now been introduced to Middles East. Currently, there is low production of jojoba globally, mainly due to the high male to female ratio in the plantations since they are mainly established from seed. Hence, a proper population stand in terms of the male: female ratio is almost impossible to maintain in the field, leading to reduced production. We report a breakthrough in the methodology for determining the sex of jojoba plants. Our research revealed that the jojoba genome contains the sequence of SRY gene, which encodes a protein similar to that found in the human Sex-Determining Region (SRY) gene. This region was amplified and matched with sequences found in papaya and human. ResultsThe jojoba-SRY regions (SRY gene) was amplified, and we closely matched with sequences of SRY in the papaya and human. The complete sequence of the DNA was depo...
Methods in molecular biology (Clifton, N.J.), 2017
Transient expression of foreign genes in plant tissue is a valuable tool for testing the efficacy... more Transient expression of foreign genes in plant tissue is a valuable tool for testing the efficacy of transformation methods. In this work, we present, for the first time, the utilization of agroinjection as an efficient transformation system for gene delivery in date palm fruit. The research utilized Agrobacterium tumefaciens strain LBA4404 harboring the binary vector pRI201-AN-GUS carrying the beta-glucuronidase (GUS) gene, under the control of a CaMV 35S and kanamycin (NPTII) as an antibiotic gene under the control of a NOS promoter. Based on histochemical assay of agroinjected fruit for the GUS gene expressions, this protocol has proved to be an efficient and reliable tool for transgene expression in date palm. PCR for plasmid DNA, extracted from the transformed Agrobacterium, demonstrated the generation of the expected amplicon, corresponding to the GUS gene using GUS primers.
Reliable in vitro regeneration protocol for tomato plant materials has optimized. Isolated and ch... more Reliable in vitro regeneration protocol for tomato plant materials has optimized. Isolated and characterized N gene from jojoba using PCR-based methods was transferred to tomato plants. The transgenic tomato plants generated bearing the jojoba N gene demonstrated that N gene confers a hypersensitive response and effectively localizes ToMV as compared with ToMV to sites of inoculation in transgenic tomato, as it also does in tobacco. The ability to reconstruct N-mediated resistance response to ToMV in tomato demonstrates the utility of using isolated resistance genes to protect crop plants from virus diseases. Furthermore, it stated that all the components necessary for N-mediated resistance conserved in tomato.
The potato (Solanum tuberosum L.) is a major food crop in Egypt. The main problem in the program ... more The potato (Solanum tuberosum L.) is a major food crop in Egypt. The main problem in the program of conventional seed potato production is the low rate of multiplication in fi eld conditions and the susceptibility of potato to diseases such as fungi, bacteria and viruses, thereby resulting in poor quality and yields. Recently, plant tissue culture technology has become very popular and has a visible impact on the production of virus free pre-basic seed potatoes. This study was aimed at producing virus free microtubers in vitro, to investigate the stimulating effects of low doses of gamma irradiation on microtuber mean number, mean fresh weight and size. Among the gamma radiation doses tested (1.5, 2, 2.5, 5 and 10 Gy), the 5 and 10 Gy doses gave the highest number of microtubers, had signifi cant effects on microtuber weight increase and also generated the highest size microtubers (180 cm 3 ). Additionally, nine potato unique markers were identifi ed among the 45 polymorphic bands, ...
Plant response to salt stress and the mechanism of salt tolerance have received major focus by pl... more Plant response to salt stress and the mechanism of salt tolerance have received major focus by plant biology researchers. Biotic stresses cause extensive losses in agricultural production globally, but abiotic stress causes significant increase in the methylglyoxal (MG) level of GlyoxalaseI (Gly I). Identification of salt-tolerant genes when characterizing their phenotypes will help to identify novel genes using polymerase chain reaction (PCR) to amplify the DNA coding region for glyoxalase I. This method is specific, requiring only genomic DNA and two pairs of PCR primers, and involving two successive PCR reactions. This method was used rapidly and easily identified glyoxalase I sequences as salt-tolerant genes from Jojoba (Simmondsia chinensis (Link) Schneider). In the present study, the glyoxalase I gene was isolated, amplified by PCR using gene-specific primers and sequenced from the jojoba plant, then compared with other glyoxalase I sequences in other plants and glyoxalase I g...
The present study aimed to identify some metabolites products obtained from different jojoba call... more The present study aimed to identify some metabolites products obtained from different jojoba callus tissue extracts using gas chromatography/mass spectrometry analysis (GC-MS). It is known that Jojoba, a medicinal and oil-yielding, has multi-purpose uses. In addition, it produces toxins, fatty acids, phenolic compounds and other secondary metabolites from callus identified using GC-MS. Despite the direct effect of 2,4-D on directing the explant towards callus induction, it has interaction effect with Kin and achieves hormonal balance inside the explant. There is a significant interaction effect between Kin and 2,4-D. The increased concentration of both Kin and 2,4-D raises the extent of nodal segments response towards proliferation, development, and callus induction. Our results showed that the interaction effect between Kin and 2,4-D is the best results for phytochemical active constituents formation achieved from callus cultured. Qualitative results revealed that increasing cytoki...
The random amplified polymorphic DNA (RAPD) technique wa s u s ed to screen markers of sex determ... more The random amplified polymorphic DNA (RAPD) technique wa s u s ed to screen markers of sex determination in this species . A 520 and 650 bp RAPD marker, marker linked t o sex determination in jojoba (male), was found in all the jojoba male but not in the female p la n t s ; it s exclus iveness to male plants . Also, A 420 bp marker linked to sex determination in jojoba (female), wa s fo u n d in a ll the jojoba fe ma le . Our data of s imilarity index reveal that there is low relationship between male and female wit h mo s t o f RAPD primers . RAPD profiles of PCR products us ing primers s ry combined wit h mo h 19 indicated that unique DNA fragmen t wit h 4 bands (1200,700, 400 and 300 bp) is present in female clones and this band is completely absent in the male clone. Howe v e r, o t h e r p olymorphic bands were detected among either male or female clones with other primers tes ted. T h e primers s ry combined with moh19 gave t h e h ighes t numbers of bands (7) and percentage o...
Reliable in vitro regeneration protocol for tomato plant materials has optimized. Isolated and ch... more Reliable in vitro regeneration protocol for tomato plant materials has optimized. Isolated and characterized N gene from jojoba using PCR-based methods was transferred to tomato plants. The transgenic tomato plants generated bearing the jojoba N gene demonstrated that N gene confers a hypersensitive response and effectively localizes ToMV as compared with ToMV to sites of inoculation in transgenic tomato, as it also does in tobacco. The ability to reconstruct N-mediated resistance response to ToMV in tomato demonstrates the utility of using isolated resistance genes to protect crop plants from virus diseases. Furthermore, it stated that all the components necessary for N-mediated resistance conserved in tomato.
ABSTRACT The effect of (IBA, IAA, and NAA) as auxins and their different concentrations on root f... more ABSTRACT The effect of (IBA, IAA, and NAA) as auxins and their different concentrations on root formation of chandler strawberry was studied. The results showed that IBA then IAA gave the highest means for all parameters and NAA significantly affect all the vegetative parameters. On the other hand, the IBA has a significant effect on root formation characteristics where No. of roots/culture (9.31a, 7.38b). The data reported also, that the optimum concentration for shoot growth was at 0.3 ppm followed by 0.6 ppm of examined auxins except for fresh and dry weight of shoot (5.44a & 1.38a g) which gave highest means with the NAA and Generally that concentration of (0,9 ppm) of examined auxins gave the lowest values. While, a medium free of any hormones (0.0 ppm) gave the best results concerning root growth and formation then 0.3 then 0.6 ppm, respectively. Our study declared that root formation and growth was better in auxins free MS medium on contrary to what was expected as for most fruit trees that need auxins to direct its explants for root formation.
Sex determination in the early developmental stages of dioecious crops is economically-beneficial... more Sex determination in the early developmental stages of dioecious crops is economically-beneficial. During this study, a human homology of SRY gene was successfully identified in dioecious crops. SRY gene sequences of date palm and jojoba were submitted to GenBank under the accession numbers KC577225 and MK991776, respectively. This is the first report regarding the novel sex-determination methodology of four dioecious plants (jojoba, date palm, papaya, and pistachios). SRY sex gene was found in all the tested dioecious plant and human samples. This novel approach is simple and of significant importance for breeders. It facilitates the unambiguous selection of jojoba and date palm female plants at an early age and reduces the plantation cost of cultivating non-productive male plants. This is a rapid sexdetermination technique for dioecious plants and mammals at an early stage. This technique specifically targets the SRY sequence that has been comprehensively investigated in humans. The kit development for the SRY-based sex determination of various crops is in progress.
BackgroundJojoba (Simmondsia Chinensis L.) is a dioecious perennial evergreen shrub native to the... more BackgroundJojoba (Simmondsia Chinensis L.) is a dioecious perennial evergreen shrub native to the south-western desert of North America and has now been introduced to Middles East. Currently, there is low production of jojoba globally, mainly due to the high male to female ratio in the plantations since they are mainly established from seed. Hence, a proper population stand in terms of the male: female ratio is almost impossible to maintain in the field, leading to reduced production. We report a breakthrough in the methodology for determining the sex of jojoba plants. Our research revealed that the jojoba genome contains the sequence of SRY gene, which encodes a protein similar to that found in the human Sex-Determining Region (SRY) gene. This region was amplified and matched with sequences found in papaya and human. ResultsThe jojoba-SRY regions (SRY gene) was amplified, and we closely matched with sequences of SRY in the papaya and human. The complete sequence of the DNA was depo...
Methods in molecular biology (Clifton, N.J.), 2017
Transient expression of foreign genes in plant tissue is a valuable tool for testing the efficacy... more Transient expression of foreign genes in plant tissue is a valuable tool for testing the efficacy of transformation methods. In this work, we present, for the first time, the utilization of agroinjection as an efficient transformation system for gene delivery in date palm fruit. The research utilized Agrobacterium tumefaciens strain LBA4404 harboring the binary vector pRI201-AN-GUS carrying the beta-glucuronidase (GUS) gene, under the control of a CaMV 35S and kanamycin (NPTII) as an antibiotic gene under the control of a NOS promoter. Based on histochemical assay of agroinjected fruit for the GUS gene expressions, this protocol has proved to be an efficient and reliable tool for transgene expression in date palm. PCR for plasmid DNA, extracted from the transformed Agrobacterium, demonstrated the generation of the expected amplicon, corresponding to the GUS gene using GUS primers.
Reliable in vitro regeneration protocol for tomato plant materials has optimized. Isolated and ch... more Reliable in vitro regeneration protocol for tomato plant materials has optimized. Isolated and characterized N gene from jojoba using PCR-based methods was transferred to tomato plants. The transgenic tomato plants generated bearing the jojoba N gene demonstrated that N gene confers a hypersensitive response and effectively localizes ToMV as compared with ToMV to sites of inoculation in transgenic tomato, as it also does in tobacco. The ability to reconstruct N-mediated resistance response to ToMV in tomato demonstrates the utility of using isolated resistance genes to protect crop plants from virus diseases. Furthermore, it stated that all the components necessary for N-mediated resistance conserved in tomato.
The potato (Solanum tuberosum L.) is a major food crop in Egypt. The main problem in the program ... more The potato (Solanum tuberosum L.) is a major food crop in Egypt. The main problem in the program of conventional seed potato production is the low rate of multiplication in fi eld conditions and the susceptibility of potato to diseases such as fungi, bacteria and viruses, thereby resulting in poor quality and yields. Recently, plant tissue culture technology has become very popular and has a visible impact on the production of virus free pre-basic seed potatoes. This study was aimed at producing virus free microtubers in vitro, to investigate the stimulating effects of low doses of gamma irradiation on microtuber mean number, mean fresh weight and size. Among the gamma radiation doses tested (1.5, 2, 2.5, 5 and 10 Gy), the 5 and 10 Gy doses gave the highest number of microtubers, had signifi cant effects on microtuber weight increase and also generated the highest size microtubers (180 cm 3 ). Additionally, nine potato unique markers were identifi ed among the 45 polymorphic bands, ...
Plant response to salt stress and the mechanism of salt tolerance have received major focus by pl... more Plant response to salt stress and the mechanism of salt tolerance have received major focus by plant biology researchers. Biotic stresses cause extensive losses in agricultural production globally, but abiotic stress causes significant increase in the methylglyoxal (MG) level of GlyoxalaseI (Gly I). Identification of salt-tolerant genes when characterizing their phenotypes will help to identify novel genes using polymerase chain reaction (PCR) to amplify the DNA coding region for glyoxalase I. This method is specific, requiring only genomic DNA and two pairs of PCR primers, and involving two successive PCR reactions. This method was used rapidly and easily identified glyoxalase I sequences as salt-tolerant genes from Jojoba (Simmondsia chinensis (Link) Schneider). In the present study, the glyoxalase I gene was isolated, amplified by PCR using gene-specific primers and sequenced from the jojoba plant, then compared with other glyoxalase I sequences in other plants and glyoxalase I g...
The present study aimed to identify some metabolites products obtained from different jojoba call... more The present study aimed to identify some metabolites products obtained from different jojoba callus tissue extracts using gas chromatography/mass spectrometry analysis (GC-MS). It is known that Jojoba, a medicinal and oil-yielding, has multi-purpose uses. In addition, it produces toxins, fatty acids, phenolic compounds and other secondary metabolites from callus identified using GC-MS. Despite the direct effect of 2,4-D on directing the explant towards callus induction, it has interaction effect with Kin and achieves hormonal balance inside the explant. There is a significant interaction effect between Kin and 2,4-D. The increased concentration of both Kin and 2,4-D raises the extent of nodal segments response towards proliferation, development, and callus induction. Our results showed that the interaction effect between Kin and 2,4-D is the best results for phytochemical active constituents formation achieved from callus cultured. Qualitative results revealed that increasing cytoki...
The random amplified polymorphic DNA (RAPD) technique wa s u s ed to screen markers of sex determ... more The random amplified polymorphic DNA (RAPD) technique wa s u s ed to screen markers of sex determination in this species . A 520 and 650 bp RAPD marker, marker linked t o sex determination in jojoba (male), was found in all the jojoba male but not in the female p la n t s ; it s exclus iveness to male plants . Also, A 420 bp marker linked to sex determination in jojoba (female), wa s fo u n d in a ll the jojoba fe ma le . Our data of s imilarity index reveal that there is low relationship between male and female wit h mo s t o f RAPD primers . RAPD profiles of PCR products us ing primers s ry combined wit h mo h 19 indicated that unique DNA fragmen t wit h 4 bands (1200,700, 400 and 300 bp) is present in female clones and this band is completely absent in the male clone. Howe v e r, o t h e r p olymorphic bands were detected among either male or female clones with other primers tes ted. T h e primers s ry combined with moh19 gave t h e h ighes t numbers of bands (7) and percentage o...
Reliable in vitro regeneration protocol for tomato plant materials has optimized. Isolated and ch... more Reliable in vitro regeneration protocol for tomato plant materials has optimized. Isolated and characterized N gene from jojoba using PCR-based methods was transferred to tomato plants. The transgenic tomato plants generated bearing the jojoba N gene demonstrated that N gene confers a hypersensitive response and effectively localizes ToMV as compared with ToMV to sites of inoculation in transgenic tomato, as it also does in tobacco. The ability to reconstruct N-mediated resistance response to ToMV in tomato demonstrates the utility of using isolated resistance genes to protect crop plants from virus diseases. Furthermore, it stated that all the components necessary for N-mediated resistance conserved in tomato.
ABSTRACT The effect of (IBA, IAA, and NAA) as auxins and their different concentrations on root f... more ABSTRACT The effect of (IBA, IAA, and NAA) as auxins and their different concentrations on root formation of chandler strawberry was studied. The results showed that IBA then IAA gave the highest means for all parameters and NAA significantly affect all the vegetative parameters. On the other hand, the IBA has a significant effect on root formation characteristics where No. of roots/culture (9.31a, 7.38b). The data reported also, that the optimum concentration for shoot growth was at 0.3 ppm followed by 0.6 ppm of examined auxins except for fresh and dry weight of shoot (5.44a & 1.38a g) which gave highest means with the NAA and Generally that concentration of (0,9 ppm) of examined auxins gave the lowest values. While, a medium free of any hormones (0.0 ppm) gave the best results concerning root growth and formation then 0.3 then 0.6 ppm, respectively. Our study declared that root formation and growth was better in auxins free MS medium on contrary to what was expected as for most fruit trees that need auxins to direct its explants for root formation.
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