Moraglia - Research

The traditional methods of genetic improvement based on breeding and selection

The use of Mutagenesis in the genetic improvement

The traditional methods of genetic improvement based on breeding and selection

The traditional methods of genetic improvement based on breeding and subsequent selection of progeny started in Moraglia Company about sixty years ago.

These methods still today remain the principal activity of the company even if the evolution of scientific knowledge about the genetic improvement of vegetable species have produced great changes, different opportunities and possibilities in breeding and selection.

The characteristics of plant and flowers of carnation that are suitable to be improved

The aim of all breeding programs is to obtain new varieties that show the amelioration of specific characters respect to the varieties used before. There are many characteristics both of the plant and of the flower that determine the success of a variety worldwide and some of them are listed in table n. 1.

Table n. 1

About the plant productivity growth speed uniformity of production stem rigidity

plant height presence of lateral buds Fusarium resistance summer or winter cultivation

About the leaves consistence dimension color

About sepals number shape consistence color

About petals shape dimension kind of edge number

consistence low temperature resistance

About flower shape dimension vase life color

creation of new colors way of flowering seasonal changes of color

About trade resistance to transport resistance to manipolation resistance to conservation

Effectively, the first step in a breeding program is the choice of the characteristics to work with and it depends on the breeder subjectivity and sensitivity, but also on the complete knowledge of carnations physiology and methods of growing, and on the proper knowledge of expectations and demands of the market.

Intuition and breeder sensitivity must be therefore joined to deep scientific, economic and commercial knowledge.
Moraglia Company obtains a twofold advantage from the fact that first creates and then trades its own varieties: the first contacts with selected growers that cultivate new varieties in different environmental conditions get useful suggestions about those characteristics that could be further on improved, while the direct contact with national and international markets produce important indications about fashions and preferences of buyers.

The technique of breeding

The technique of breeding could be schematically subdivided in four subsequent moments:

a) the choice of parental lines
The choice of parental lines is perhaps the most delicate question using the traditional methods of breeding and selection. In fact they have to be chose and evaluate according to the objectives that have been pointed out in the program of genetic improvement. It is possible to chose parental lines within the varieties that show the characters that have to be improved or within those varieties that even if don’t show these characters could improve as well the qualitative aspects of an existing variety. Furthermore it’s important to evaluate the genetic characteristic of each character: some of those that are shown in table n. 1 are dominant, others are recessive, some are monogenic and others polygenic.
Thanks to the long-term experience developed in the art of breeding and selection, nowadays Moraglia Company owns a large collection of female and male line appropriately selected for specific characters; this collection is constantly enriched and up-dated to satisfy the continuous market’s demands of novelties.

b) the growing of parental line
Female and male lines are grown in standard conditions from May to December in sterilezed benches on a peat-perlite substrate. The environment will be sunny and windy.

c) the technique of hybridisation
- Collection of pollen: the first moment of cross is the collection of pollen on the varieties that have been selected as male lines. The anthers with pollen are collected inside a Petri dish when they are dehiscent during the hottest hours of daytime. Pollen could be conserved up to three days at the temperature of 1-3°C.
– Preparation of female line: on female plant it is necessary to remove first lateral flowers and buds and then calyx, corolla and stamens leaving only the pistil with the stigma well developed.
- Pollination: it takes place during the hottest hours about midday in sunny and moderate windy days. The stem of female plant is gently bended on Petri dish to bring into contact the stigma with pollen. If stigma is at the fully ripeness it retains pollen.

d) the collection of seeds
The capsules that have been fecundated ripe in 30-60 days depending on climatic conditions and the specific characteristic of the varieties that have been used. The seeds are harvested when they appeared completely black and they are preserved until sowing in a dry and dark place.

Selection of Progeny

Seed are sown in a seed-bed on a proper substrate with constant temperature and humidity conditions. During germination is important to pay attention to temperature and humidity conditions to avoid the death of the young plants owing to thermal or hydric stresses or in consequence of collar (neck) diseases.
Young plants are transplanted after 30-40 days depending on climatic conditions; leaves and roots might be enough developed to ensure plant to get over transplanting stress, too. Plants are then grown following the traditional technique.
Selection of plants starts after 3 months: the way and the speed of growth, the habitus of plant and all the vegetative characteristics are valuated earlier because they strictly determine the success of cultivation; in a second time the characteristics of flower are estimated.
After two years of selection in our nursery the most promising varieties are tested for Fusarium resistance and at the third year they are finally grown and evaluated under different environmental conditions in Italy and in other European countries in order to verify the uniformity and the stability of the new variety under different pedologic and climatic conditions.

The use of Mutagenesis in the genetic improvement

Clonal selection

The clonal selection allows to choose within a variety those individuals that completely express the specific characters of that variety and to eliminate the ones that show unfavourable characteristics occasionally arose in consequence of spontaneous mutagenesis.

Selection of “sport”

The “sport” are individuals that present spontaneous genetic mutations that often induce changes in the colour of the flower or in other characteristics of plant. If these alterations are positive they are selected and preserved.

The induced mutagenesis

Some plants of well known and appreciate varieties are grown in vitro and are exposed to chemical and physical agents that induce genetic mutations. These alterations produce changes in the colour, in the speed of growth, in the margin of petals, ecc. and could be successively selected.
Disease resistant lines and clones with different speed of growth have been obtained by somaclonal variation and in vitro selection techniques.

Table n. 1
About the plant	productivity	growth speed	uniformity of production	stem rigidity
	plant height	presence of lateral buds	Fusarium resistance	summer or winter cultivation
About the leaves	consistence	dimension	color
About sepals	number	shape	consistence	color
About petals	shape	dimension	kind of edge	number
	consistence	low temperature resistance
About flower	shape	dimension	vase life	color
	creation of new colors	way of flowering	seasonal changes of color
About trade	resistance to transport	resistance to manipolation	resistance to conservation

	Effectively, the first step in a breeding program is the choice of the characteristics to work with and it depends on the breeder subjectivity and sensitivity, but also on the complete knowledge of carnations physiology and methods of growing, and on the proper knowledge of expectations and demands of the market. Intuition and breeder sensitivity must be therefore joined to deep scientific, economic and commercial knowledge. Moraglia Company obtains a twofold advantage from the fact that first creates and then trades its own varieties: the first contacts with selected growers that cultivate new varieties in different environmental conditions get useful suggestions about those characteristics that could be further on improved, while the direct contact with national and international markets produce important indications about fashions and preferences of buyers.
	The technique of breeding The technique of breeding could be schematically subdivided in four subsequent moments: a) the choice of parental lines The choice of parental lines is perhaps the most delicate question using the traditional methods of breeding and selection. In fact they have to be chose and evaluate according to the objectives that have been pointed out in the program of genetic improvement. It is possible to chose parental lines within the varieties that show the characters that have to be improved or within those varieties that even if don’t show these characters could improve as well the qualitative aspects of an existing variety. Furthermore it’s important to evaluate the genetic characteristic of each character: some of those that are shown in table n. 1 are dominant, others are recessive, some are monogenic and others polygenic. Thanks to the long-term experience developed in the art of breeding and selection, nowadays Moraglia Company owns a large collection of female and male line appropriately selected for specific characters; this collection is constantly enriched and up-dated to satisfy the continuous market’s demands of novelties. b) the growing of parental line Female and male lines are grown in standard conditions from May to December in sterilezed benches on a peat-perlite substrate. The environment will be sunny and windy.
		c) the technique of hybridisation - Collection of pollen: the first moment of cross is the collection of pollen on the varieties that have been selected as male lines. The anthers with pollen are collected inside a Petri dish when they are dehiscent during the hottest hours of daytime. Pollen could be conserved up to three days at the temperature of 1-3°C. – Preparation of female line: on female plant it is necessary to remove first lateral flowers and buds and then calyx, corolla and stamens leaving only the pistil with the stigma well developed. - Pollination: it takes place during the hottest hours about midday in sunny and moderate windy days. The stem of female plant is gently bended on Petri dish to bring into contact the stigma with pollen. If stigma is at the fully ripeness it retains pollen.

	d) the collection of seeds The capsules that have been fecundated ripe in 30-60 days depending on climatic conditions and the specific characteristic of the varieties that have been used. The seeds are harvested when they appeared completely black and they are preserved until sowing in a dry and dark place. Selection of Progeny Seed are sown in a seed-bed on a proper substrate with constant temperature and humidity conditions. During germination is important to pay attention to temperature and humidity conditions to avoid the death of the young plants owing to thermal or hydric stresses or in consequence of collar (neck) diseases. Young plants are transplanted after 30-40 days depending on climatic conditions; leaves and roots might be enough developed to ensure plant to get over transplanting stress, too. Plants are then grown following the traditional technique. Selection of plants starts after 3 months: the way and the speed of growth, the habitus of plant and all the vegetative characteristics are valuated earlier because they strictly determine the success of cultivation; in a second time the characteristics of flower are estimated. After two years of selection in our nursery the most promising varieties are tested for Fusarium resistance and at the third year they are finally grown and evaluated under different environmental conditions in Italy and in other European countries in order to verify the uniformity and the stability of the new variety under different pedologic and climatic conditions. The use of Mutagenesis in the genetic improvement
	Clonal selection The clonal selection allows to choose within a variety those individuals that completely express the specific characters of that variety and to eliminate the ones that show unfavourable characteristics occasionally arose in consequence of spontaneous mutagenesis. Selection of “sport” The “sport” are individuals that present spontaneous genetic mutations that often induce changes in the colour of the flower or in other characteristics of plant. If these alterations are positive they are selected and preserved.
	The induced mutagenesis Some plants of well known and appreciate varieties are grown in vitro and are exposed to chemical and physical agents that induce genetic mutations. These alterations produce changes in the colour, in the speed of growth, in the margin of petals, ecc. and could be successively selected. Disease resistant lines and clones with different speed of growth have been obtained by somaclonal variation and in vitro selection techniques.