Method of ex vitro sowing, germination, growth and conversion of plant somatic embryos or germinants, and nutrient medium used therefor

Abstract

A method of sowing a somatic plant embryo or germinant of a conifer species to facilitate subsequent development of the embryo or germinant into an autotrophic seedling. The method involves the following steps carried out ex vitro in non-sterile conditions: providing a nutrient medium comprising particles of a solid component present within a flowable or semi-solid component containing water and a carbohydrate nutrient for the embryo or germinant, dispensing a quantity of the nutrient medium onto a surface of a porous solid growth substrate for the somatic plant embryo or germinant, and contacting the plant embryo or germinant with the nutrient medium. The nutrient medium has a fluidity such that at least some of the flowable or semi-solid component containing the carbohydrate nutrient remains in contact with the embryo or germinant at least until the embryo or germinant establishes vigorous growth under environmental conditions effective for such growth. The particles of the solid component are adapted to remain in contact with the embryo or germinant after of the flowable or semi-solid material dissipates, thereby providing continuing physical support for the embryo or germinant after the dissipation. The invention also relates to seedlings produced in this way and to the nutrient solution.

Description

FIELD OF THE INVENTION [0001] This invention relates to the sowing, germination, growth, and conversion of somatic embryos or germinants of conifer species. More particularly, the invention relates to sowing, and development of somatic embryos or germinants ex vitro in a porous solid growth substrate, such as soil or peat, to allow the embryos or germinants to develop into autotrophic seedlings and subsequently into mature plants in non-sterile conditions. BACKGROUND OF THE INVENTION [0002] Dependence on the use of zygotic seed in breeding programs, where success relies on seed availability, particularly that of genetically superior seeds, often leads to low returns on investment. The situation is exacerbated in tree breeding and improvement programs especially where conifer species are used, because the time span from flower bud initiation to seed maturation is usually one to three years. Climatic events and pest infection contribute to seed production variability from year to year...

AI Technical Summary

Benefits of technology

"The present invention provides a method for producing complete, independent, autotrophic plants from conifer somatic embryos or germinants in conventional nursery conditions. This is achieved by providing a nutrient medium containing particles of a solid component that remains in contact with the embryo or germinant after the flowable or semi-solid component has dissipated. The nutrient medium can be dispensed onto a porous solid growth substrate and can contain a variety of ingredients including water, simple organic and inorganic plant nutrients, protective agents, and gelling agents. The method involves exposing the embryo or germinant to environmental conditions and allowing it to develop into an autotrophic seedling. The invention also provides a porous solid growth substrate for the embryo or germinant, which can be pre-wetted and charged with conventional fertilizers. The nutrient medium can be formulated in either sterile or non-sterile conditions and can be dispensed onto a porous solid growth substrate using hand or machinery. The conifer somatic embryos can be placed in sterile or non-sterile conditions and the sown containers can be placed in sterile or non-sterile conditions to allow the plant somatic embryos or germinants to grow into complete, independent, autotrophic plants."

Problems solved by technology

Dependence on the use of zygotic seed in breeding programs, where success relies on seed availability, particularly that of genetically superior seeds, often leads to low returns on investment.

The primary challenge in utilising somatic embryogenesis for clonal forestry in plantation programs is the development of cost effective and scaleable methods of somatic embryo culture to produce autotrophic and acclimatised seedlings.

Second, after the somatic embryo has germinated and grown under sterile conditions, the young seedling is transplanted into conventional nursery growing systems.

No approaches are taught or otherwise disclosed in the prior art for sowing encapsulated somatic embryos and / or artificial seed and / or manufactured seed into conventional growing systems using conventional sowing equipment.

There are many disadvantages associated with in vitro protocols.

The most significant are: 1) the repeated manual handling of each individual embryo in the germination and transplanting steps; 2) the stringent requirement for sterile techniques and culture conditions through all steps until somatic germinants are transplanted out of the in vitro germination environment into horticultural growing media; and 3) the difficulty in acclimatizing in vitro plantlets into ex vitro nursery environments.

Therefore, the art of traditional in vitro protocols has an inherent nature of low efficiency and high cost, characteristics that are prohibitive to mass production of somatic seedlings.

These undesirable characteristics make the commercial production of somatic seedlings less competitive than that of the zygotic seedlings (Sutton and Polonenko 1999).

However, no commercial equipment currently exists which can reliably, aseptically, and cost-effectively perform the in vitro protocols for germination and gorwht of naked somatic embryos and subsequent transplanting of seedlings into conventional propagation systems (Roberts et al., 1995; reviews by Sakamoto et al., 1995; and Sutton and Polonenko 1999).

There are also numerous biological and operational disadvantages inherent in using gel-encapsulated somatic embryos.

Biologically, the most significant disadvantage is the much lower germination vigour and conversion success into plants than corresponding zygotic seeds, as seen in the prior art protocols for encapsulating or otherwise coating somatic embryos (Redenbaugh et al., 1993; Carlson & Hartle, 1995; Gray et al., 1995).

Because such a “culture medium is highly susceptible to invasion by phytopathogens, which can result in death or retard the growth of the embryos”, they teach that the containers, substrate, nutrient solutions and other components of their system must be biologically sterile.

Even though the art taught by such methods may be practised to produce somatic seedlings, such methods are labour-intensive and bear characteristics of low efficiency, high cost and impracticability for mass production of somatic seedlings in a nursery environment.

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