Diploid and haploid embryogenesis in Larixleptolepis, L. decidua, and their reciprocal hybrids

1990 ◽  
Vol 20 (1) ◽  
pp. 9-14 ◽  
Author(s):  
P. von Aderkas ◽  
K. Klimaszewska ◽  
J. M. Bonga

Diploid and haploid embryogenesis was induced in two Larix species (L. decidua and L. leptolepis) and their reciprocal hybrids. Diploid embryogenic tissue was initiated in immature zygotic embryos isolated with the micropylar half of the megagametophyte left attached. These were placed either on modified LM or MSG medium supplemented with the growth regulators 2,4-D and 6-benzyladenine. MSG medium was solidified with either gellan gum or agar. There was no appreciable difference in response between the two. Haploid embryogenesis was induced in isolated megagametophytes placed on modified LM medium supplemented with 2,4-D and 6-benzyladenine. Diploid embryogenic tissue was subcultured on medium with growth regulators, but haploid embryogenic tissue grew well on medium without growth regulators. There were few morphological differences between the diploid and haploid embryogenic tissue. In all species and hybrids, haploid cultures contained more coenocytic long cells. Binucleate cells were most common, but tetranucleate and octanucleate cells were also present. Haploid cultures showed poorer organization than the diploid ones, with only a few cultures having well-developed embryoids. Haploid tissue originated from expanded cells of the megagametophyte. Diploid tissue originated from the suspensor region of the zygotic embryo; it proliferated from isolated clusters of meristematic cells in early embryoids. Diploid and haploid cultures differed not only from the outset, but also in the mature embryoids they produced.


2007 ◽  
Vol 59 (3) ◽  
pp. 199-202 ◽  
Author(s):  
Dragana Stojicic ◽  
Branka Uzelac ◽  
Dusica Janosevic ◽  
Ljubinka Culafic ◽  
Snezana Budimir

The potential for somatic embryogenesis in zygotic embryo and megagametophyte cultures of Pinus heldreichii was examined. Somatic embryogenesis was initiated from megagametophytes containing immature zygotic embryos at early stages of development. An induction frequency of up to 6.7% was obtained on Gresshoff and Doy medium in the presence of 2 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5 mg/l benzyladenine (BA). Formation and further proliferation of embryogenic tissue were achieved upon transfer of explants to a medium with reduced levels of growth regulators. Somatic embryos are being cultured for further development. .



2009 ◽  
Vol 39 (8) ◽  
pp. 1566-1574 ◽  
Author(s):  
Cathy L. Hargreaves ◽  
Cathie B. Reeves ◽  
Jens I. Find ◽  
Keiko Gough ◽  
Puthiyaparambil Josekutty ◽  
...  

The principal aim of this investigation was to improve somatic embryogenesis initiation and to enhance representation of families and genotypes within those families of Pinus radiata D. Don. A total of 19 open-pollinated seed families, many with unrelated and weakly related parents, were tested. Optimum stage of cone maturity for initiation success was tested by five collections made at 1 week intervals, spanning the developmental period from pro-embryo to cotyledonary embryos. Two media were compared; embryo-development media (EDM6) and a modified Litvay medium (Glitz). Two zygotic embryo explant-preparation techniques were tested; embryos with retained megagametophytes and excised embryos. Proliferating embryogenic tissues were obtained from all four treatments (2850 explants per treatment, 570 per collection time) for the 19 families. The best initiation rates were achieved with a combination of Glitz medium with excised zygotic embryos, with 55% of explants from all collections and all families combined giving rise to proliferating embryogenic tissue. At the optimal collection time for each of the families, this treatment gave a range of 47%–97% initiation success with an average of 70% per family.



1990 ◽  
Vol 20 (6) ◽  
pp. 810-817 ◽  
Author(s):  
M. R. Becwar ◽  
R. Nagmani ◽  
S. R. Wann

Immature zygotic embryo explants (isolated or with intact megagametophytes) from 10 loblolly pine (Pinustaeda L.) clones (7-34, 7-56, 11-9, 11-16, 11-25, 10-1003, 10-1007, 10-1011, 10-1018, and 10-1019) were surveyed for their potential to form embryogenic tissue from the suspensor region of zygotic embryos. After over 14 000 explants were cultured, embryogenic cultures were initiated from explants of 8 of the 10 clones; only explants from clones 11-25 and 10-1019 were not responsive. Embryogenic tissue was initiated from zygotic embryos with intact megagametophytes on MSG basal medium with no exogenous plant growth regulators or with 2–5 mg/L 2,4-dichlorophenoxy acetic acid (2,4-D) and 0–1 mg/L N6-benzyladenine (BA). The highest initiation frequency (5%) was obtained from isolated zygotic embryos of clone 7-34 less than 0.5 mm in length just prior to cotyledon primordia development on DCR basal medium with 3 mg/L 2,4-D and 0.5 mg/L BA. Two types of embryogenic cultures were maintained on medium with 2,4-D and BA: (i) those that contained pre-embryonal masses of cells interspersed with unaggregated suspensorlike cells, but which rarely contained well-formed somatic embryos, and (ii) those that frequently contained well-formed somatic embryos. Somatic embryo development from both types of cultures progressed to a precotyledonary stage on medium with 2.6 mg/L abscisic acid.



1993 ◽  
Vol 23 (5) ◽  
pp. 873-876 ◽  
Author(s):  
R. Nagmani ◽  
A.M. Diner ◽  
G.C. Sharma

Isolated zygotic embryos and female gametophytes containing zygotic embryos were cultured on MSG and DCR basal media, supplemented with three different carbon sources added individually to the medium at four levels each. The media also contained various levels of 2,4-dichlorophenoxy acetic acid (2,4-D) and N6-benzyladenine (BA). Embryogenic tissue extruded from female gametophytes during 4 weeks in culture on media containing either glucose or maltose or sucrose. Embryogenic tissue initiation was most frequently from explants collected on July 14, 1992, when the zygotic embryos within the female gametophytes were precotyledonary. A total of 33 embryogenic cultures were initiated from 944 explants cultured. One of 192 explants cultured on basal media with no growth regulators produced embryogenic tissue. The embryogenic tissue showed numerous somatic embryos at stages 1 and 2 of development, corresponding to their zygotic embryo counterparts.



2015 ◽  
Vol 122 (3) ◽  
pp. 629-638 ◽  
Author(s):  
Darley Aparecido Tavares Ferreira ◽  
Mariana Cansian Sattler ◽  
Carlos Roberto Carvalho ◽  
Wellington Ronildo Clarindo


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