Genome Size and Genetic Homogeneity of Regenerated Plants: Methods and Applications

Cellular and Molecular Tools for the Investigation of Somatic Embryogenesis in Medicago Species

Author(s): Anelia V. Iantcheva * .

Pp: 105-124 (20)

DOI: 10.2174/9789815165555123010008

* (Excluding Mailing and Handling)

Abstract

The chapter presents the knowledge accumulated on the recent investigation of somatic embryogenesis (SE) in genera Medicago. The role of 2,4-D in the process of induction of embryogenic potential in diploid Medicago and its transport by the combined action of auxin transporters or diffusion of dissociated molecules is discussed. Among the many methods for studying the process, this chapter is focused on cellular and molecular tools – flow cytometry, assessment of expression level of SE related transcripts of key genes of auxin inducible process and different PCR techniques. Our recent studies on the process of SE in M. truncatula are focused on the role of the two genes MtLAX3 (an auxin transmembrane transporter) and a transcriptional factor MtARF-B3 (an auxin response factor, containing a B3-binding domain). The transcription profiles of these genes are evaluated and their expression patterns are assessed during indirect somatic embryogenesis – steps of callus formation, embryogenic zone formation and the stages of globular, torpedo and cotyledonary embryos. The localization of expression during the process of SE is traced by the β-glucuronidase reporter gene (GUS) under the control of the promoters of these genes. Inverse PCR (IPCR) and Transposon display (TD) are techniques which evaluate transposition and new retrotransposon copies in the investigated mutant lines, and we used these methods as markers for the efficiency of the induction phase of the process of SE. The use of all these methods turns light on a better understanding of the process of somatic embryogenesis in the model species Medicago truncatula and other annual medics.

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      • 16. vendor/laravel/framework/src/Illuminate/Routing/ControllerDispatcher.php:43
      • 17. vendor/laravel/framework/src/Illuminate/Routing/Route.php:260
    • (select j.user_access_id,j.volume_id,j.user_id,j.from_ip,j.to_ip,j.type,j.track,"Trial Access" as access_base_text, CASE j.type WHEN 111 THEN true ELSE false END as terms_popup, "S" as content_download_type, null as restricted_user_access_key, null as restricted_user_access_type, CASE j.track WHEN 0 THEN false ELSE true END as tracksAccessByVolume,"chapter" as access_level, j.chapter_id from `user_access_chapter_trail` as `j` inner join `user_access_info` as `info` on `j`.`user_access_id` = `info`.`user_access_id` where ((j.from_ip <= 51368230 and j.to_ip >=51368230 )) and ((j.from_date <= 1739834833 and j.to_date >= 1739834833 and j.perpetual= 0) or (j.from_date is null and j.to_date is null and j.perpetual= 1)) and j.chapter_id = 20217) union (select j.user_access_id,j.volume_id,j.user_id,j.from_ip,j.to_ip,j.type,j.track,"Subscribed" as access_base_text, false as terms_popup, "S" as content_download_type, null as restricted_user_access_key, null as restricted_user_access_type, CASE j.track WHEN 0 THEN false ELSE true END as tracksAccessByVolume,"chapter" as access_level, j.chapter_id from `user_access_chapter_corporate` as `j` inner join `user_access_info` as `info` on `j`.`user_access_id` = `info`.`user_access_id` where ((j.from_ip <= 51368230 and j.to_ip >=51368230 )) and ((j.from_date <= 1739834833 and j.to_date >= 1739834833 and j.perpetual= 0) or (j.from_date is null and j.to_date is null and j.perpetual= 1)) and j.chapter_id = 20217) union (select j.user_access_id,j.volume_id,j.user_id,j.from_ip,j.to_ip,j.type,j.track,"Token Based Access" as access_base_text, false as terms_popup, "R" as content_download_type, j.uc_id as restricted_user_access_key, "v" as restricted_user_access_type, CASE j.track WHEN 0 THEN false ELSE true END as tracksAccessByVolume,"chapter" as access_level, j.chapter_id from `user_access_chapter_token` as `j` inner join `user_access_info` as `info` on `j`.`user_access_id` = `info`.`user_access_id` where ((j.from_ip <= 51368230 and j.to_ip >=51368230 )) and ((j.from_date <= 1739834833 and j.to_date >= 1739834833 and j.perpetual= 0) or (j.from_date is null and j.to_date is null and j.perpetual= 1)) and j.chapter_id = 20217)
      900μsalphaeurekaselec_live_10_06_2022UserAccess.php#1266
      Backtrace
      • 13. app/Models/UserAccess/UserAccess.php:1266
      • 14. app/Http/Controllers/ChapterController.php:1549
      • 15. vendor/laravel/framework/src/Illuminate/Routing/Controller.php:54
      • 16. vendor/laravel/framework/src/Illuminate/Routing/ControllerDispatcher.php:43
      • 17. vendor/laravel/framework/src/Illuminate/Routing/Route.php:260
    • select b.banner_link, b.banner_name, b.banner_script, b.banner_remarks, b.banner_file_name, b.banner_placement, bd.content_type, bd.content_id from `tbl_banner` as `b` inner join `tbl_banner_detail` as `bd` on `bd`.`banner_id` = `b`.`banner_id` inner join `users` as `u` on `u`.`id` = `b`.`updated_by` where `b`.`banner_ebook` = 1 and `bd`.`content_type` = 'B' and `bd`.`content_id` = 3571 and `b`.`banner_from_date` <= 1739834833 and `b`.`banner_to_date` >= 1739834833 and `b`.`banner_status` = 'A' order by `b`.`banner_id` desc
      1.56msalphaeurekaselec_live_10_06_2022Banner.php#95
      Bindings
      • 0: 1
      • 1: B
      • 2: 3571
      • 3: 1739834833
      • 4: 1739834833
      • 5: A
      Backtrace
      • 13. app/Models/Banner.php:95
      • 14. app/Http/helpers.php:404
      • 17. vendor/laravel/framework/src/Illuminate/Filesystem/Filesystem.php:124
      • 18. vendor/laravel/framework/src/Illuminate/View/Engines/PhpEngine.php:58
      • 19. vendor/laravel/framework/src/Illuminate/View/Engines/CompilerEngine.php:73
    • select count(*) as aggregate from `uc_cart_products` where cart_id='588565c4b9dae2dff3b4d37752e787d6'
      250μsalphaeurekaselec_live_10_06_2022Cart.php#271
      Backtrace
      • 15. app/Models/Cart.php:271
      • 16. view::layouts._header:260
      • 18. vendor/laravel/framework/src/Illuminate/Filesystem/Filesystem.php:124
      • 19. vendor/laravel/framework/src/Illuminate/View/Engines/PhpEngine.php:58
      • 20. vendor/laravel/framework/src/Illuminate/View/Engines/CompilerEngine.php:73
    App\Models\Chapter
    1Chapter.php#?
    App\Models\Ebook
    1Ebook.php#?
        _token
        6R9MHFAdygMLWnXd3rIeIq4VDBhClWfksv8fXbl9
        uc_cart_id
        588565c4b9dae2dff3b4d37752e787d6
        _previous
        array:1 [ "url" => "http://alpha.eurekaselect.com/chapter/20217" ]
        _flash
        array:2 [ "old" => [] "new" => [] ]
        PHPDEBUGBAR_STACK_DATA
        []
        path_info
        /chapter/20217
        status_code
        200
        
        status_text
        OK
        format
        html
        content_type
        text/html; charset=UTF-8
        request_query
        []
        
        request_request
        []
        
        request_headers
        0 of 0
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        request_cookies
        0 of 0
        array:1 [ "wayfindertarget" => null ]
        response_headers
        0 of 0
        array:5 [ "content-type" => array:1 [ 0 => "text/html; charset=UTF-8" ] "cache-control" => array:1 [ 0 => "no-cache, private" ] "date" => array:1 [ 0 => "Mon, 17 Feb 2025 23:27:13 GMT" ] "set-cookie" => array:2 [ 0 => "XSRF-TOKEN=eyJpdiI6InB1VytQTEpBL1RJU1BZNXI2ZUMrblE9PSIsInZhbHVlIjoib0NrbWZaOFZLdUR5MDBsckp1cGJ4S0dBZzV4dHNuNGVwZEZxME92b1VUMEpFUEFmblRqSllXOG9kWHNkRDNtS0xVVzZxcjI4SW5tSytPc1dzQUZDcHFNU0pLeCsvV0dJRlZVQ1NWZ3Z6TVhSOGVZU0RleURoU1NWS1d3VDVia00iLCJtYWMiOiI2MTkxMTM1MWU3NGE0ODNlYmY3ZjExMTE5MmIyNjlkZTg4ODkzMmI2ZDlkYmQzNmYwZmU0ZDEzOTA2ZThhMDlhIiwidGFnIjoiIn0%3D; expires=Thu, 20 Feb 2025 11:27:13 GMT; Max-Age=216000; path=/; secure; samesite=laxXSRF-TOKEN=eyJpdiI6InB1VytQTEpBL1RJU1BZNXI2ZUMrblE9PSIsInZhbHVlIjoib0NrbWZaOFZLdUR5MDBsckp1cGJ4S0dBZzV4dHNuNGVwZEZxME92b1VUMEpFUEFmblRqSllXOG9kWHNkRDNtS0xVVzZxc" 1 => "alphaeurekaselectnet_session=eyJpdiI6InZMc1VOUE9xcXhqa2E2aEpCb2NSdmc9PSIsInZhbHVlIjoiQXJnT2JBdE9aZElyYnFPRGJPR1RIRDF4UnorT0dFLzZjcklSMkdCRHMya0F0c2pUbDRjYVQwbm5nUGZycThNV0RTU0s2KzhrQ3pKUG1Ta2I4bzh1L0NieFJYM0NFWDhWdndtL2VFZXZEb25jV3RKU0FiajFISzJTdHVGOFVKS3AiLCJtYWMiOiI5MTAxODUzOTUzNjJhY2MyNWRlYWVhYjhiZTZkOTA4MmI4YmM3ZTQzNjQ3YTcxNjZhNGRhMDEyNzcxZjZjNTFlIiwidGFnIjoiIn0%3D; path=/; secure; httponly; samesite=laxalphaeurekaselectnet_session=eyJpdiI6InZMc1VOUE9xcXhqa2E2aEpCb2NSdmc9PSIsInZhbHVlIjoiQXJnT2JBdE9aZElyYnFPRGJPR1RIRDF4UnorT0dFLzZjcklSMkdCRHMya0F0c2pUbDRjYVQwbm5" ] "Set-Cookie" => array:2 [ 0 => "XSRF-TOKEN=eyJpdiI6InB1VytQTEpBL1RJU1BZNXI2ZUMrblE9PSIsInZhbHVlIjoib0NrbWZaOFZLdUR5MDBsckp1cGJ4S0dBZzV4dHNuNGVwZEZxME92b1VUMEpFUEFmblRqSllXOG9kWHNkRDNtS0xVVzZxcjI4SW5tSytPc1dzQUZDcHFNU0pLeCsvV0dJRlZVQ1NWZ3Z6TVhSOGVZU0RleURoU1NWS1d3VDVia00iLCJtYWMiOiI2MTkxMTM1MWU3NGE0ODNlYmY3ZjExMTE5MmIyNjlkZTg4ODkzMmI2ZDlkYmQzNmYwZmU0ZDEzOTA2ZThhMDlhIiwidGFnIjoiIn0%3D; expires=Thu, 20-Feb-2025 11:27:13 GMT; path=/; secureXSRF-TOKEN=eyJpdiI6InB1VytQTEpBL1RJU1BZNXI2ZUMrblE9PSIsInZhbHVlIjoib0NrbWZaOFZLdUR5MDBsckp1cGJ4S0dBZzV4dHNuNGVwZEZxME92b1VUMEpFUEFmblRqSllXOG9kWHNkRDNtS0xVVzZxc" 1 => "alphaeurekaselectnet_session=eyJpdiI6InZMc1VOUE9xcXhqa2E2aEpCb2NSdmc9PSIsInZhbHVlIjoiQXJnT2JBdE9aZElyYnFPRGJPR1RIRDF4UnorT0dFLzZjcklSMkdCRHMya0F0c2pUbDRjYVQwbm5nUGZycThNV0RTU0s2KzhrQ3pKUG1Ta2I4bzh1L0NieFJYM0NFWDhWdndtL2VFZXZEb25jV3RKU0FiajFISzJTdHVGOFVKS3AiLCJtYWMiOiI5MTAxODUzOTUzNjJhY2MyNWRlYWVhYjhiZTZkOTA4MmI4YmM3ZTQzNjQ3YTcxNjZhNGRhMDEyNzcxZjZjNTFlIiwidGFnIjoiIn0%3D; path=/; secure; httponlyalphaeurekaselectnet_session=eyJpdiI6InZMc1VOUE9xcXhqa2E2aEpCb2NSdmc9PSIsInZhbHVlIjoiQXJnT2JBdE9aZElyYnFPRGJPR1RIRDF4UnorT0dFLzZjcklSMkdCRHMya0F0c2pUbDRjYVQwbm5" ] ]
        session_attributes
        0 of 0
        array:5 [ "_token" => "6R9MHFAdygMLWnXd3rIeIq4VDBhClWfksv8fXbl9" "uc_cart_id" => "588565c4b9dae2dff3b4d37752e787d6" "_previous" => array:1 [ "url" => "http://alpha.eurekaselect.com/chapter/20217" ] "_flash" => array:2 [ "old" => [] "new" => [] ] "PHPDEBUGBAR_STACK_DATA" => [] ]
        ClearShow all
        Date ↕MethodURLData
        #12025-02-17 23:27:13GET/chapter/20217112192