The pollen genome of Echinacea purpurea ‘Magnus’ (purple coneflower) was amplified using a modifi... more The pollen genome of Echinacea purpurea ‘Magnus’ (purple coneflower) was amplified using a modified primer extension pre-amplification (PEP) procedures followed by AFLP (amplified fragment length polymorphism) analyses of individual pollen grains. Twenty-four AFLP primer pairs were identified that produced 104 scorable markers in eleven linkage groups. This is the first report on the production of AFLP markers from individual pollen grains for genetic mapping in coneflower. Coneflowers are important to both the ornamental and medicinal herb industries.
This is a PDF file of an article that has undergone enhancements after acceptance, such as the ad... more This is a PDF file of an article that has undergone enhancements after acceptance, such as the addition of a cover page and metadata, and formatting for readability, but it is not yet the definitive version of record. This version will undergo additional copyediting, typesetting and review before it is published in its final form, but we are providing this version to give early visibility of the article. Please note that, during the production process, errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
As an important crop for US agricultural security and sustainable agriculture, cotton genetic imp... more As an important crop for US agricultural security and sustainable agriculture, cotton genetic improvement requires development of innovative analytical tools. Genetic analyses based on individual gametes provide several advantages including minimum sample requirement, genetic identification of male parent and overcoming the complexity of polyploidy due to haploid nature of microspores. This research used tetraploid cotton (G. hirsutum x G. barbadense) chromosome substitution (CS) lines with G. barbadense chromosomes 17 and 25 in G. hirsutum background. The individually isolated pollen grains were germinated to release their DNAs and genomic DNA was increased by modified primer extension pre-amplification (PEP) using MasterAmp™ Extra-Long PCR Kit (EPICENTRE®, Madison, WI). Also from select cotton lines, microspores just released from tetrad developmental stage were individually isolated and gamete DNA was extracted as well as amplified through multiple displacement amplification (MDA) using REPLI-g Single Cell Kit (QIAGEN, Valencia, CA). The parental samples along with PEP and MDA amplified individual gamete DNAs were then analyzed using simple sequence repeat (SSR) as well as IRD-800 and IRD-700 labeled (Li-Cor, Lincoln, NE) amplified fragment length polymorphism (AFLP) methods. Nineteen SSR and 28 AFLP primer pairs were used to analyze TM-1, 3-79, CS-B17 and CS-B25 cotton lines. The amplification of parental SSR and AFLP markers from both mature pollen as well as early free microspore samples provided unique tools for comparative genetic studies using agarose and polyacrylamide gel electrophoresis, respectively.
Cotton (Gossypium spp.) is the world's third most important oilseed crop as well as an economic s... more Cotton (Gossypium spp.) is the world's third most important oilseed crop as well as an economic source of protein. However, the presence of (-)-gossypol limits the nutritional value of cottonseeds. To genetically profile relevant traits, seeds of the two cotton parental lines TM-1 (G. hirstum) and 3-79 (G. barbadense) along with their 17 chromosomal substitution (CS) progenies were quantified for (-)-and (+)-enantiomer levels. The TM-1 parent was found to contain a higher ratio of (+)-:(-)-gossypol enantiomers (13.34:8.52 µg/mg) compared to that of 3-79 levels (6.69:7.41 µg/mg) in the seeds. Among progeny CS-B (containing substituted chromosomes from G. barbadense in G. hirsutum background) lines, only CS-B17 was found to contain significantly lower (7.65 µg/mg) level of (+)-enantiomer in its seeds, and also had the lowest (5.09 µg/mg) amount of (-)-gossypol among all lines. A singular protocol was then devised for RNA yields from polysaccharide-rich cottonseeds of CS-B17 and its parents. Transcription derived fragments (TDFs) analyses of these three lines were conducted on seeds harvested at 28, 35 and 42 days post anthesis (DPA). 64 TDFs profiles were analyzed using differential expression analyses, and similarities were revealed between 3-79 and CS-B17 along with unique expression trends for CS-B17. Such expression analyses, which are based on single chromosome differences with TM-1 parent while reflecting seed nutritional merit of 3-79 parent, may facilitate cotton breeding endeavors.
Dry pollen of diploid Solanum clone 9507-04, Cotton chromosomal substitution line and Echinacea p... more Dry pollen of diploid Solanum clone 9507-04, Cotton chromosomal substitution line and Echinacea purpurea "Magnus" were isolated and their cells contents released in individual tubes containing water or pollen germination media, respectively.Six polymorphic markers from individual pollen grains of the diploid potato were amplified using two random amplified polymorphic DNA (RAPD) primers, i.e., UBC291 and UBC504. The procedures for primer extension pre-amplification (PEP) for genome and subsequent amplified fragment length polymorphism (AFLP) analysis with 13 primers pairs were tested on the upland cotton line. The cotton parental AFLP markers were scored for their presence or absences in the 35 pollen samples and majority of the markers were found monomorphic as expected from the gametes of a near isogenic line.PEP and AFLP procedures for linkage mapping were standardized usingpurpleconeflower pollen grains. From 64 possible AFLP primer pairs,37.5% amplified 104 markers for coneflowerthat were linked in 11 linkage groupsranging from 13 to 134 centimorgans (cM). The 409 cM sizedlinkage map constructed forE. purpurea "Magnus"genome had average distance of 2.4 cM to 22 cMfor the two to 55 polymorphic markers identifiedwithin the 11 groups.The protocols described in this report would serve to enhance breeding efficiencies in plantsimportant for food,fiber, ornamental and medicinal industries.
Potato is fourth most important crop in the world, and anther culture is used towards its genetic... more Potato is fourth most important crop in the world, and anther culture is used towards its genetic improvements. Anther culture, as a breeding tool, can influence leaf shape related characters of this species, which warrants further studies. Two potato clones were compared through their anther-derived plants based on five characteristics: overall shape of terminal or primary leaflet, shape of leaflet's tip, shape of leaflet's bases, amount of leaf anthocyanin, and pubescence. The anther-derived plants of clone 9507-04 had been regenerated from calli/embryos while clone 6028-02 produced plantlets directly from the anthers. Also, 46-94% of anther-derived plants from clone 6028-02 were similar to the parent while only 37-60% of offspring were found to be parental kinds for clone 9507-04. On confirmation of the DNA contents and/or ploidy of anther-derived plants, it was found that potato clones 9507-04 and 6028-02 produced 77% and 100% monoploids, respectively. The variations, in androgenic fitness as well as culture-response among potato anther-donors, may have an important role for ploidy and leaf morphology differences among anther-derived siblings. By identifying these variations among potato clones, the findings in this report have subsequent applications for generating androgenic plants, with the aim of improving light capture, water balance, as well as temperature regulations by the leaves of crop plants.
A diploid Solanum clone 9507-04 was used to grow anther-derived plantlets that were putatively mo... more A diploid Solanum clone 9507-04 was used to grow anther-derived plantlets that were putatively monoploid. Sixteen RAPD primers were used to observe the RAPD profiles from anther-donor and anther-derived materials. A total of 56 RAPD markers were scored and each marker was identified according to the primer's designation and size (kilo base pairs) of the amplified fragment. Of 56 RAPD markers assayed, 44.6% did not follow the expected 1:1 Me ndelian segregation (χ 2 >4.26, P<0.05) in the anther-derived progeny. The selected six morphological traits also segregated in anther-derived plants including the nine that retained 3-4 of the parental leaf characters. The 12 distortion-causing RAPD markers (χ2=8.89-17.64, P<0.003) with above 84% presence in the progeny were retained in all above nine plants. Such preference of parental markers in potato anther-derived progeny may indicate a linkage with androgenesis.
Energy is lifeline in development and progress of a country, with pivotal use in industrial and a... more Energy is lifeline in development and progress of a country, with pivotal use in industrial and agricultural sectors. While fossil fuels are primary source of energy, its rapid depletion is major concerns of the world today. Developed countries are finding alternates to strengthen its economic backbone by resolving limited supply of energy issues and meeting their enormous demands. Plants can offer better alternative as biofuels to address the energy crisis and meet projected energy demand. Recently, sorghum based research remained center of attention due to its exceptional properties to grow under dry, and hot environmental conditions with limited water requirement. All parts of sorghum have economic values with usage in syrup, sugar, fuel, alcohol, bedding and paper production. Whereas, Sorghum stalks are enriched source of carbohydrates with 16-18% of fermentable sugar that makes it potential candidate for bioethanol production. Bioethanol is considered environmental friendly as it reduces greenhouse gases and replaces MTBE (Methyl tert-butyl ether) pollutants in air. Yet, the benefits of sorghum as biofuels comes with a challenge of rapid degradation of sugar, therefore immediate harvest after maturity can ensure high sugar content. This review covers scope and recent research on sorghum in Pakistan and indicates its usage as an ideal feedstock to meet present energy crisis. Currently, developed countries are exploiting sorghum in bioethanol production due to its high tolerance to drought and salt, and improved sugar content in lieu of using sugarcane and maize. Moreover, studies involving high-tech research and role of microRNA in high yield and improved sugar content in biofuel production of sorghum are also addressed in this review.
This paper is based on a project that evaluated the benefits of biotechnology training workshops ... more This paper is based on a project that evaluated the benefits of biotechnology training workshops for its integration into school curriculum that is dependent on teachers' knowledge, their teaching confidence, as well as their opinion towards biotechnology. Teachers, as well as a parallel group comprising of college students, were given pre-and post-training tests to assess changes in their knowledge and opinion about biotechnology. Participant teachers' confidence in teaching biotechnology was also assessed. These evaluations revealed a significant increase in the post-test scores compared to the pre-test for teachers (df = 22, t= 12.706, p<0.0001) and college students (df= 12, t= 5.584, p<0.0001). In addition, the teachers (df= 39, t= 8.078, p<0.0001) as well as the college students (df= 7, t= 4.174, p=0.0042) registered a significant increase in their positive attitude towards biotechnology after the training workshops. Biotechnology teaching confidence of the participant educators also showed a significant increase (df= 35, t= 6.393, p<.0001) after the workshops. However, urban teachers had a significantly greater (df= 34, t=-3.191, p=0.0030) gain of 0.376 in their post-training teaching confidence compared to the 0.145 gain by their rural counterparts. Additionally, a statistically significant (r= 0.868, r-squared= 0.7534, p= .0219) relationship was found between teaching confidence and positive opinion of the teachers regarding biotechnology. Since teachers' pedagogical approach and adoption of biotechnology curriculum is dependent on the above issues, results presented in this study are important for designing future biotechnology training workshops.
This manuscript has been reproduced from the microfilm master. UMI films the text directly from t... more This manuscript has been reproduced from the microfilm master. UMI films the text directly from the original or copy submitted. Thus, some thesis and dissertation copies are in typewriter face, while others may be frorn any type of computer printer. ...
Daylily (Hemerocallis sp. ‘Stella de Oro’) callus cultures initiated from ovules were bombarded w... more Daylily (Hemerocallis sp. ‘Stella de Oro’) callus cultures initiated from ovules were bombarded with gold particles coated with plasmid harboring Basta® resistance gene. Resulting putative transgenic calli were selected after 3 wk on semi-solid Murashige and Skoog’s (MS) basal medium supplemented with 10 mg L-1 1-naphthaleneacetic acid, 2 mg L-1 6-benzylaminopurine and 3 mg L-1 phosphinothricin (PPT). Surviving calli regenerated shoots after 2 mo on semi-solid MS medium supplemented with 2 mg L-1 thiadiazuron and 1 mg L-1 PPT. Polymerase chain reaction and Southern blotting were used to confirm independent transformation events. Key words: Basta® resistance, in vitro, Hemerocallis
... Key words: β-glucuronidase, Basta® resistance, in vitro, Ulmus parvifolia Aziz, AN, Sauvé, RJ... more ... Key words: β-glucuronidase, Basta® resistance, in vitro, Ulmus parvifolia Aziz, AN, Sauvé, RJ, Zhou, S. et Meng, X. 2003. Transformation ... 1987) and stable transformation (Klein et al. 1988; Sanford 1990) in other plant types. This ...
The pollen genome of Echinacea purpurea ‘Magnus’ (purple coneflower) was amplified using a modifi... more The pollen genome of Echinacea purpurea ‘Magnus’ (purple coneflower) was amplified using a modified primer extension pre-amplification (PEP) procedures followed by AFLP (amplified fragment length polymorphism) analyses of individual pollen grains. Twenty-four AFLP primer pairs were identified that produced 104 scorable markers in eleven linkage groups. This is the first report on the production of AFLP markers from individual pollen grains for genetic mapping in coneflower. Coneflowers are important to both the ornamental and medicinal herb industries.
This is a PDF file of an article that has undergone enhancements after acceptance, such as the ad... more This is a PDF file of an article that has undergone enhancements after acceptance, such as the addition of a cover page and metadata, and formatting for readability, but it is not yet the definitive version of record. This version will undergo additional copyediting, typesetting and review before it is published in its final form, but we are providing this version to give early visibility of the article. Please note that, during the production process, errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
As an important crop for US agricultural security and sustainable agriculture, cotton genetic imp... more As an important crop for US agricultural security and sustainable agriculture, cotton genetic improvement requires development of innovative analytical tools. Genetic analyses based on individual gametes provide several advantages including minimum sample requirement, genetic identification of male parent and overcoming the complexity of polyploidy due to haploid nature of microspores. This research used tetraploid cotton (G. hirsutum x G. barbadense) chromosome substitution (CS) lines with G. barbadense chromosomes 17 and 25 in G. hirsutum background. The individually isolated pollen grains were germinated to release their DNAs and genomic DNA was increased by modified primer extension pre-amplification (PEP) using MasterAmp™ Extra-Long PCR Kit (EPICENTRE®, Madison, WI). Also from select cotton lines, microspores just released from tetrad developmental stage were individually isolated and gamete DNA was extracted as well as amplified through multiple displacement amplification (MDA) using REPLI-g Single Cell Kit (QIAGEN, Valencia, CA). The parental samples along with PEP and MDA amplified individual gamete DNAs were then analyzed using simple sequence repeat (SSR) as well as IRD-800 and IRD-700 labeled (Li-Cor, Lincoln, NE) amplified fragment length polymorphism (AFLP) methods. Nineteen SSR and 28 AFLP primer pairs were used to analyze TM-1, 3-79, CS-B17 and CS-B25 cotton lines. The amplification of parental SSR and AFLP markers from both mature pollen as well as early free microspore samples provided unique tools for comparative genetic studies using agarose and polyacrylamide gel electrophoresis, respectively.
Cotton (Gossypium spp.) is the world's third most important oilseed crop as well as an economic s... more Cotton (Gossypium spp.) is the world's third most important oilseed crop as well as an economic source of protein. However, the presence of (-)-gossypol limits the nutritional value of cottonseeds. To genetically profile relevant traits, seeds of the two cotton parental lines TM-1 (G. hirstum) and 3-79 (G. barbadense) along with their 17 chromosomal substitution (CS) progenies were quantified for (-)-and (+)-enantiomer levels. The TM-1 parent was found to contain a higher ratio of (+)-:(-)-gossypol enantiomers (13.34:8.52 µg/mg) compared to that of 3-79 levels (6.69:7.41 µg/mg) in the seeds. Among progeny CS-B (containing substituted chromosomes from G. barbadense in G. hirsutum background) lines, only CS-B17 was found to contain significantly lower (7.65 µg/mg) level of (+)-enantiomer in its seeds, and also had the lowest (5.09 µg/mg) amount of (-)-gossypol among all lines. A singular protocol was then devised for RNA yields from polysaccharide-rich cottonseeds of CS-B17 and its parents. Transcription derived fragments (TDFs) analyses of these three lines were conducted on seeds harvested at 28, 35 and 42 days post anthesis (DPA). 64 TDFs profiles were analyzed using differential expression analyses, and similarities were revealed between 3-79 and CS-B17 along with unique expression trends for CS-B17. Such expression analyses, which are based on single chromosome differences with TM-1 parent while reflecting seed nutritional merit of 3-79 parent, may facilitate cotton breeding endeavors.
Dry pollen of diploid Solanum clone 9507-04, Cotton chromosomal substitution line and Echinacea p... more Dry pollen of diploid Solanum clone 9507-04, Cotton chromosomal substitution line and Echinacea purpurea "Magnus" were isolated and their cells contents released in individual tubes containing water or pollen germination media, respectively.Six polymorphic markers from individual pollen grains of the diploid potato were amplified using two random amplified polymorphic DNA (RAPD) primers, i.e., UBC291 and UBC504. The procedures for primer extension pre-amplification (PEP) for genome and subsequent amplified fragment length polymorphism (AFLP) analysis with 13 primers pairs were tested on the upland cotton line. The cotton parental AFLP markers were scored for their presence or absences in the 35 pollen samples and majority of the markers were found monomorphic as expected from the gametes of a near isogenic line.PEP and AFLP procedures for linkage mapping were standardized usingpurpleconeflower pollen grains. From 64 possible AFLP primer pairs,37.5% amplified 104 markers for coneflowerthat were linked in 11 linkage groupsranging from 13 to 134 centimorgans (cM). The 409 cM sizedlinkage map constructed forE. purpurea "Magnus"genome had average distance of 2.4 cM to 22 cMfor the two to 55 polymorphic markers identifiedwithin the 11 groups.The protocols described in this report would serve to enhance breeding efficiencies in plantsimportant for food,fiber, ornamental and medicinal industries.
Potato is fourth most important crop in the world, and anther culture is used towards its genetic... more Potato is fourth most important crop in the world, and anther culture is used towards its genetic improvements. Anther culture, as a breeding tool, can influence leaf shape related characters of this species, which warrants further studies. Two potato clones were compared through their anther-derived plants based on five characteristics: overall shape of terminal or primary leaflet, shape of leaflet's tip, shape of leaflet's bases, amount of leaf anthocyanin, and pubescence. The anther-derived plants of clone 9507-04 had been regenerated from calli/embryos while clone 6028-02 produced plantlets directly from the anthers. Also, 46-94% of anther-derived plants from clone 6028-02 were similar to the parent while only 37-60% of offspring were found to be parental kinds for clone 9507-04. On confirmation of the DNA contents and/or ploidy of anther-derived plants, it was found that potato clones 9507-04 and 6028-02 produced 77% and 100% monoploids, respectively. The variations, in androgenic fitness as well as culture-response among potato anther-donors, may have an important role for ploidy and leaf morphology differences among anther-derived siblings. By identifying these variations among potato clones, the findings in this report have subsequent applications for generating androgenic plants, with the aim of improving light capture, water balance, as well as temperature regulations by the leaves of crop plants.
A diploid Solanum clone 9507-04 was used to grow anther-derived plantlets that were putatively mo... more A diploid Solanum clone 9507-04 was used to grow anther-derived plantlets that were putatively monoploid. Sixteen RAPD primers were used to observe the RAPD profiles from anther-donor and anther-derived materials. A total of 56 RAPD markers were scored and each marker was identified according to the primer's designation and size (kilo base pairs) of the amplified fragment. Of 56 RAPD markers assayed, 44.6% did not follow the expected 1:1 Me ndelian segregation (χ 2 >4.26, P<0.05) in the anther-derived progeny. The selected six morphological traits also segregated in anther-derived plants including the nine that retained 3-4 of the parental leaf characters. The 12 distortion-causing RAPD markers (χ2=8.89-17.64, P<0.003) with above 84% presence in the progeny were retained in all above nine plants. Such preference of parental markers in potato anther-derived progeny may indicate a linkage with androgenesis.
Energy is lifeline in development and progress of a country, with pivotal use in industrial and a... more Energy is lifeline in development and progress of a country, with pivotal use in industrial and agricultural sectors. While fossil fuels are primary source of energy, its rapid depletion is major concerns of the world today. Developed countries are finding alternates to strengthen its economic backbone by resolving limited supply of energy issues and meeting their enormous demands. Plants can offer better alternative as biofuels to address the energy crisis and meet projected energy demand. Recently, sorghum based research remained center of attention due to its exceptional properties to grow under dry, and hot environmental conditions with limited water requirement. All parts of sorghum have economic values with usage in syrup, sugar, fuel, alcohol, bedding and paper production. Whereas, Sorghum stalks are enriched source of carbohydrates with 16-18% of fermentable sugar that makes it potential candidate for bioethanol production. Bioethanol is considered environmental friendly as it reduces greenhouse gases and replaces MTBE (Methyl tert-butyl ether) pollutants in air. Yet, the benefits of sorghum as biofuels comes with a challenge of rapid degradation of sugar, therefore immediate harvest after maturity can ensure high sugar content. This review covers scope and recent research on sorghum in Pakistan and indicates its usage as an ideal feedstock to meet present energy crisis. Currently, developed countries are exploiting sorghum in bioethanol production due to its high tolerance to drought and salt, and improved sugar content in lieu of using sugarcane and maize. Moreover, studies involving high-tech research and role of microRNA in high yield and improved sugar content in biofuel production of sorghum are also addressed in this review.
This paper is based on a project that evaluated the benefits of biotechnology training workshops ... more This paper is based on a project that evaluated the benefits of biotechnology training workshops for its integration into school curriculum that is dependent on teachers' knowledge, their teaching confidence, as well as their opinion towards biotechnology. Teachers, as well as a parallel group comprising of college students, were given pre-and post-training tests to assess changes in their knowledge and opinion about biotechnology. Participant teachers' confidence in teaching biotechnology was also assessed. These evaluations revealed a significant increase in the post-test scores compared to the pre-test for teachers (df = 22, t= 12.706, p<0.0001) and college students (df= 12, t= 5.584, p<0.0001). In addition, the teachers (df= 39, t= 8.078, p<0.0001) as well as the college students (df= 7, t= 4.174, p=0.0042) registered a significant increase in their positive attitude towards biotechnology after the training workshops. Biotechnology teaching confidence of the participant educators also showed a significant increase (df= 35, t= 6.393, p<.0001) after the workshops. However, urban teachers had a significantly greater (df= 34, t=-3.191, p=0.0030) gain of 0.376 in their post-training teaching confidence compared to the 0.145 gain by their rural counterparts. Additionally, a statistically significant (r= 0.868, r-squared= 0.7534, p= .0219) relationship was found between teaching confidence and positive opinion of the teachers regarding biotechnology. Since teachers' pedagogical approach and adoption of biotechnology curriculum is dependent on the above issues, results presented in this study are important for designing future biotechnology training workshops.
This manuscript has been reproduced from the microfilm master. UMI films the text directly from t... more This manuscript has been reproduced from the microfilm master. UMI films the text directly from the original or copy submitted. Thus, some thesis and dissertation copies are in typewriter face, while others may be frorn any type of computer printer. ...
Daylily (Hemerocallis sp. ‘Stella de Oro’) callus cultures initiated from ovules were bombarded w... more Daylily (Hemerocallis sp. ‘Stella de Oro’) callus cultures initiated from ovules were bombarded with gold particles coated with plasmid harboring Basta® resistance gene. Resulting putative transgenic calli were selected after 3 wk on semi-solid Murashige and Skoog’s (MS) basal medium supplemented with 10 mg L-1 1-naphthaleneacetic acid, 2 mg L-1 6-benzylaminopurine and 3 mg L-1 phosphinothricin (PPT). Surviving calli regenerated shoots after 2 mo on semi-solid MS medium supplemented with 2 mg L-1 thiadiazuron and 1 mg L-1 PPT. Polymerase chain reaction and Southern blotting were used to confirm independent transformation events. Key words: Basta® resistance, in vitro, Hemerocallis
... Key words: β-glucuronidase, Basta® resistance, in vitro, Ulmus parvifolia Aziz, AN, Sauvé, RJ... more ... Key words: β-glucuronidase, Basta® resistance, in vitro, Ulmus parvifolia Aziz, AN, Sauvé, RJ, Zhou, S. et Meng, X. 2003. Transformation ... 1987) and stable transformation (Klein et al. 1988; Sanford 1990) in other plant types. This ...
Uploads
Papers by Ahmad Aziz