An ultrastructural study on the early development of Zea mays somatic embryos

1991 ◽  
Vol 69 (4) ◽  
pp. 858-865 ◽  
Author(s):  
P. F. Fransz ◽  
J. H. N. Schel

Friable embryogenic callus, obtained from immature embryos of Zea mays L., was cultured on N6 medium supplemented with 1 mg/L 2,4-dichlorophenoxyacetic acid, 6 mM proline, and 2% sucrose. Cultured tissue fragments containing several globular embryoids were excised and examined by light and electron microscopy to follow the early development of maize embryoids. The somatic embryos consist of an apical region and a suspensor region. Cells of the apical region are small, cytoplasm rich, and mitotically active. They contain much starch and numerous bundles of microtubules. Suspensor cells are larger and more vacuolated. A high metabolic activity in both cell types is indicated by the presence of many organelles, coated vesicles, and multivesicular bodies. Transition units appear to form intermediate stages between the embryogenic callus cells and the somatic embryo. A transition unit consists of a group of embryogenic cells and shows an apical and a basal region. The unit has many intercellular spaces, and within the cells areas with organelle-free cytosol are frequently observed. Key words: somatic embryogenesis, in vitro culture, ultrastructure, Zea mays L.

2016 ◽  
Vol 76 (1) ◽  
Author(s):  
Pauline D KASI ◽  
. SUMARYONO

Summary Embryogenic callus of sago (Metroxylon sagu Rottb.) has been grown on three systems of in vitro culture i.e. agar-solidified medium, liquid medium, and temporary immersion system (TIS) medium to observe and compare the development of embryogenic callus over one passage of six weeks.  A-half gram of embryogenic callus was cultured on a modified MS medium containing 10 mg/L   2,4-D and 0.1 mg/L kinetin. For histological studies, embryogenic callus was fixed in FAA and embedded in paraplast wax. Serial sections were stained with safranin 1% and observed microscopically. By the end of culture period, the development of embryogenic callus in TIS medium was relatively better than those of the other two media.  Fresh weight of callus in liquid medium and TIS increased by 6.5-fold, while on agar-solidified medium increased by 5.4-fold in six weeks.  About 40% of callus in liquid medium and TIS and 20% of callus on agar solidified medium have changed into somatic embryos at globular stage. Histology structure of embryogenic callus of the three systems of in vitro culture shows different pattern. On agar-solidified medium, secondary callus and friable embryogenic callus that consist of meristematic cells were formed. In contrast, more embryogenic cells were formed in liquid medium and TIS to support maturation process to somatic embryos. Therefore, temporary immersion system and liquid medium are recommended for maturation of embryogenic callus, whereas agar-solidified medium is for proliferation of embryogenic callus of sago.  Ringkasan Kalus embriogenik sagu (Metroxylon sagu Rottb.) telah ditumbuhkan pada tiga sistem kultur in vitro yaitu medium padat, medium cair, dan medium dengan sistem perendaman sesaat (SPS) untuk mempelajari dan mem-bandingkan perkembangan dari kalus embrio-genik selama periode enam minggu. Setengah gram kalus embriogenik dikulturkan pada medium MS modifikasi yang mengandung  2,4-D 10 mg/L dan kinetin 0,1 mg/L.  Untuk studi histologi, kalus embriogenik difiksasi dengan FAA dan embedding menggunakan lilin paraplast. Irisan diwarnai dengan safranin 1% dan diamati menggunakan mikroskop. Pada akhir periode kultur, pertumbuhan kalus pada medium dengan SPS lebih baik dibandingkan dengan medium cair dan padat. Bobot basah kalus pada  medium cair dan SPS meningkat 6,5 kali sedangkan pada medium padat meningkat 5,4 kali dalam waktu enam minggu. Sebanyak 40% kalus pada medium cair dan SPS serta 20% kalus pada medium padat berubah menjadi embrio somatik fase globuler. Struktur histologi kalus embriogenik pada ketiga jenis sistem kultur in vitro menunjukkan pola yang berbeda. Pada medium padat terjadi pembentukan kalus sekunder dan kalus embriogenik remah yang terdiri atas sel-sel meristematik. Sebaliknya pada medium cair dan SPS pembentukan sel embriogenik lebih banyak yang menunjang proses pendewasaan menjadi embrio somatik. Oleh karena itu, medium cair dan SPS direkomendasikan untuk pendewasaan kalus embriogenik, sedangkan medium padat untuk proliferasi kalus embriogenik sagu. 


2016 ◽  
Vol 76 (1) ◽  
Author(s):  
Pauline D KASI ◽  
. SUMARYONO

Summary Embryogenic callus of sago (Metroxylon sagu Rottb.) has been grown on three systems of in vitro culture i.e. agar-solidified medium, liquid medium, and temporary immersion system (TIS) medium to observe and compare the development of embryogenic callus over one passage of six weeks.  A-half gram of embryogenic callus was cultured on a modified MS medium containing 10 mg/L   2,4-D and 0.1 mg/L kinetin. For histological studies, embryogenic callus was fixed in FAA and embedded in paraplast wax. Serial sections were stained with safranin 1% and observed microscopically. By the end of culture period, the development of embryogenic callus in TIS medium was relatively better than those of the other two media.  Fresh weight of callus in liquid medium and TIS increased by 6.5-fold, while on agar-solidified medium increased by 5.4-fold in six weeks.  About 40% of callus in liquid medium and TIS and 20% of callus on agar solidified medium have changed into somatic embryos at globular stage. Histology structure of embryogenic callus of the three systems of in vitro culture shows different pattern. On agar-solidified medium, secondary callus and friable embryogenic callus that consist of meristematic cells were formed. In contrast, more embryogenic cells were formed in liquid medium and TIS to support maturation process to somatic embryos. Therefore, temporary immersion system and liquid medium are recommended for maturation of embryogenic callus, whereas agar-solidified medium is for proliferation of embryogenic callus of sago.  Ringkasan Kalus embriogenik sagu (Metroxylon sagu Rottb.) telah ditumbuhkan pada tiga sistem kultur in vitro yaitu medium padat, medium cair, dan medium dengan sistem perendaman sesaat (SPS) untuk mempelajari dan mem-bandingkan perkembangan dari kalus embrio-genik selama periode enam minggu. Setengah gram kalus embriogenik dikulturkan pada medium MS modifikasi yang mengandung  2,4-D 10 mg/L dan kinetin 0,1 mg/L.  Untuk studi histologi, kalus embriogenik difiksasi dengan FAA dan embedding menggunakan lilin paraplast. Irisan diwarnai dengan safranin 1% dan diamati menggunakan mikroskop. Pada akhir periode kultur, pertumbuhan kalus pada medium dengan SPS lebih baik dibandingkan dengan medium cair dan padat. Bobot basah kalus pada  medium cair dan SPS meningkat 6,5 kali sedangkan pada medium padat meningkat 5,4 kali dalam waktu enam minggu. Sebanyak 40% kalus pada medium cair dan SPS serta 20% kalus pada medium padat berubah menjadi embrio somatik fase globuler. Struktur histologi kalus embriogenik pada ketiga jenis sistem kultur in vitro menunjukkan pola yang berbeda. Pada medium padat terjadi pembentukan kalus sekunder dan kalus embriogenik remah yang terdiri atas sel-sel meristematik. Sebaliknya pada medium cair dan SPS pembentukan sel embriogenik lebih banyak yang menunjang proses pendewasaan menjadi embrio somatik. Oleh karena itu, medium cair dan SPS direkomendasikan untuk pendewasaan kalus embriogenik, sedangkan medium padat untuk proliferasi kalus embriogenik sagu. 


1985 ◽  
Vol 146 (3) ◽  
pp. 327-334 ◽  
Author(s):  
Kathryn K. Kamo ◽  
Michael R. Becwar ◽  
Thomas K. Hodges

Biologia ◽  
2021 ◽  
Author(s):  
Zuzana Lukacova ◽  
Denis Liska ◽  
Boris Bokor ◽  
Renata Svubova ◽  
Alexander Lux
Keyword(s):  
Zea Mays ◽  

2017 ◽  
Vol 17 (1) ◽  
pp. 9
Author(s):  
Yosi Zendra Joni ◽  
Riry Prihatini ◽  
Darda Efendi ◽  
Ika Roostika

<p>Somatic embryogenesis is a technique for regenerating embryos derived from somatic cells of various plant species. This technique along with the utilization of plant growth regulator (PGR) might benefit for mass propagation and improvement of plant species through biotechnological tools. The study aimed to determine the effect of different plant growth regu-lators, namely 6-benzyladenine (BA) and thidiazuron (TDZ) on the embryogenic callus induction as well as casein hydrolysate and malt extract on the somatic embryo development of mangosteen. The explants used were in vitro young stems of mangosteen clone Leuwiliang. This study consisted of two experiments, namely induction of embryogenic callus and formation of somatic embryo. The first experiment was arranged as factorial in a completely randomized design with BA (0 and 0.7 mg l-1) as the first factor and TDZ (0, 0.1, 0.5 and 1.0 mg l-1) as the second factor. The second experiment consisted of four treatments, i.e. casein hydrolysate and malt extract at the rate of 500 and 1,000 mg l-1. The results showed that the best medium for embryogenic callus induction was MS supplemented with 0.1 mg l-1 TDZ, which resulted semifriable calli. Casein hydrolysate and malt extract could not induce the formation of somatic embryos. After two times subcultures on the same MS medium supplemented with 0.5 mg l-1 TDZ and 0.7 mg l-1 BA, a total of 33.8 somatic embryos per explant was induced. The successful somatic embryogenesis would support mangosteen breeding and in vitro mass propagation program.</p>


2000 ◽  
Vol 12 (2) ◽  
pp. 146-155 ◽  
Author(s):  
TEREZINHA RANGEL CAMARA ◽  
LILIA WILLADINO ◽  
JOSEP MARIA TORNÉ ◽  
ALBERT MANICK ◽  
MARIA ASUNCIÓN SANTOS
Keyword(s):  
Zea Mays ◽  

O presente trabalho teve por objetivo avaliar o efeito de diferentes concentrações de NaCl (0, 68, 137 e 205 mM) sobre o cultivo in vitro de calos de dois genótipos (W64Ao2 e Arizona 8601) de milho (Zea mays L.), em meio de cultura N6, suplementado ou não com a adição de 6,0 mM de prolina. Os tratamentos consistiram da combinação dos quatro níveis de NaCl com os dois níveis de prolina, num fatorial 4x2. Os tratamentos foram mantidos durante 60 dias, com subcultivos a cada 20 dias. A taxa de crescimento dos calos e o conteúdo endógeno de putrescina, espermina, espermidina, prolina e outros aminoácidos livres foram avaliados no final do período experimental. A adição de prolina exógena favoreceu o crescimento dos calos no tratamento-controle e minorou os efeitos deletérios do estresse salino em ambos os genótipos, no nível de sal mais elevado (250mM de NaCl). A taxa de crescimento dos calos de W64Ao2 foi superior àquela dos calos de Arizona 8601, possivelmente por causa, em parte, do maior acúmulo de prolina endógena e da manutenção ativa do metabolismo das poliaminas naqueles calos.


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