Intraspecific Hybrids and Heterosis Breeding

Intraspecific hybridisation is widely used in the genetic improvement of Salvia species. The most important achievements in the case of clary sage are reported here. The hybridisation work of Elnir et al. (1991b) is of great importance first of all from a theoretical point of view. Wild cultivated chemotypes of S. sclarea originally from Israel and Russia which differed in oil composition were crossed to study the inheri tance pattern and to gain new oil composition. The essential oil distilled from the Israeli type had a citral-like odour, mentioned as a citral chemotype while the plant material of Russian origin produced the commercial type of oil. Reciprocal hybridis ation of the chemotypes resulted in two different types of hybrids, which shows the role of maternal effects. One of the hybrid groups accumulated linalool, geranyl acetate and geraniol in a relatively high amount, 22.0%, 22.5% and 22% respectively, while in the other one the presence of linalyl acetate and geranial was relatively high (12.6% and 7.2%). Besides, most of the components of the essential oil of the hybrids were intermediate between those of the parents. However, it seems, that in this case the existing intraspecific chemical variability can not be used effectively for the establishment of special cultivars, because the hybrids proved to be completely sterile, despite the observed normal meiosis (Elnir et al., 1991a).

Considerable practical results were achieved however by hybridisation of clary sage lines in Ukraine and Bulgaria. To get cultivars of high productivity, hybridisation was started in the early sixties simultaneously in both countries. In the Bulgarian breeding program the local cultivars were hybridised with the best Ukrainian types to get progenies of high productivity and cold resistance (Ilieva, 1980). As a result of the long process of selection four cultivars were developed: "Lazour", "Slunchev luch", "Roza" and "Zarya". All these cultivars surpassed the standard populations in essential oil content (which ranged between 0.20-0.24%) and in oil yield. The new cultivars were distinguishable by morphological characteristics and the time and duration of anthesis.

In the Ukraine (Zobenko et al., 1989, Bugayenko et al., 1995) the main aim of breeding S. sclarea was to get varieties of high productivity, populations owing different developmental cycles (annual and biennial) and plant material with resistance again root rot (Thielaviopsis basicola (Bek. et Br. Ferraris). By intraspecific hybridisation (between cultivars) using an effective castration method new clary sage cultivars, the annual "Crimsky odnoletniy" and "Crimsky ultraskorospely" charac terised by a very short vegetation period were bred.

In the above mentioned two countries, the effectiveness of breeding could be increased even by application of heterosis breeding. It was proven, that by means of inbreeding, uniform and highly productive lines could be developed (Goncharyuk et al., 1988). Inbreeding depression is affected by the genotype of the source variety and is, in several cases, not considerable. By continuous inbreeding Vlasova (1986) produced lines differing in both morphological and production-biological characters from the original cultivar. Even the accumulation level of essential oil increased up to 2%. Beside F1 hybrid production, these lines proved to be useful for other breeding methods, such as simple selection or synthetic varieties. Inbreeding combined with multiple selection recently led to practical results. A new late ripeness cultivar of clary sage under the name "Crimsky pozdniy" was created and introduced into large scale cultivation (Zobenko, 1995).

In Salvia species, castration is not an economically acceptable method for the effective heterosis breeding. In large scale seed production there is a need for male sterile counterparts. It seems, that in the case of S. sclarea this cannot be a problem. The occurrence of male sterility had been found—in accordance with S. nemorosa and S. officinalis (Mohan and Kans, 1990), which may contribute to the effectiveness of the hybridisation. Male sterile plants were found in Bulgaria, too (Dascalova, 1996), in a collection of freely pollinated lines and among individuals of the "Boyana" cultivar. By studying the biology of male sterility it has been established that male sterility increases the quantity of seed production compared with the seed set ratio of the fertile plants by an average 20%, due to sterilisation processes in the different stages of megagametogenesis and embryogenesis. It may even increase the value of using male-sterile plants in the heterosis breeding.

In Ukraine, cytoplasmically male sterile analogues of the breeding lines have been developed and are used for seed production (Arinstejn et al., 1985; Kovalev et al., 1988; Kovalev, 1989). It was established, that the ratio of the pollen donor parent should be at least 17-25% compared to the male-sterile mother plants. However, both in the production of hybrid seeds and in maintenance of the lines, the pollina tion will be complete only in the presence of bee families.

There has been much less experience in the intraspecific hybridization of the other Salvia species when compared to S. sclarea, as discussed above. However, the results achieved by the co-operation of Italian-Austrian groups on S. officinalis are worth a mention (Franz, 1994; Bezzi, 1996; Landi and Bertone, 1996).

In order to achieve successful breeding and cultivar development and by realising the lack of knowledge in this context, studies on the inheritance of the most impor tant plant traits by crossings, were carried out (Franz, 1994). It is obvious from the data presented in Table 6. that crossing of cineol type (P132) and a-thujone type (P191) plants resulted in both a-thujone and E-thujone type individuals, as well. Also it is a very interesting phenomenon, that while both parents contain relatively large

Table 6 Compositional segregation of hybrids made by crossing of different S. officinalis chemotypes (Franz 1994).


Main constituents of the essential oil (%)





















132 x 191.14







132 x 191.13







132 x 191.19







132 x 191.15







132x 191.15







amounts of a-thujone (29.66-34.66%), its amount decreased to below 1% in one group of hybrids. The same unexpected trend of changes can be observed in the case of the majority of the other compounds. Based on the results of the above experiment, it was admitted by the author, that additive or epistatic gene effects should be present in the regulation of formation of essential oil compounds.

Figure 2 Accumulation level of essential oil in the original population of Salvia sclarea and cv. "Trakiia" as well as in the mutant line number 40-3, in a three year experiment (Mehraz et al. 1988).

In the case of this species, instead of heterosis breeding, establishment of synthetic varieties seemed to be more advantageous, due to the biological background and high costs of Fl hybrid production. In practice, the polycross method can be easily applied, considering the good vegetative propagation possibilities of S. officinalis. For cultivar development, testing the combining ability in diallele trials, selection of lines of proper homogenity, large leaf area, considerable essential accumulation level and high productivity was carried out. The results of this work and the establishment of a synthetic variety in polycross blocks from these clones were reported by Bezzi (1996) as well as by Landi and Bertone (1996).


It is only in the last ten years, that it has been proven that the induction of mutant and polyploid forms may contribute to the improvement of the genetic background of S. sclarea. The construction of these mutant lines being carried out by colchicine treatment.

Based on the results of Savchenko (1990), the induced polyploids can be well characterised and selected from the basic population according to an index. This index is calculated by taking into account the leaf shape, its margin, colour, surface and the number of germination pores in the pollen grains just before flowering time.

The mutation breeding produced even practical results. Mekhraz et al. (1988) constructed a new cultivar producing mutants by a 24 h treatment with colchicine. The growth, development and essential oil content of mutant lines were studied and compared with both the standard cv. "Trakiia" and the initial population. It was found that the new lines had faster growth and blooming rates. Moreover, some of the lines (especially line number 40-3) produced much higher total fresh mass and essential oil yield. The accumulation level of essential oils in this special line mentioned under the name "Boyana" proved to be superior to the standards in three year experiments (Fig. 2). The essential oil composition of the selected cultivar is characterised mainly by the presence of linalyl acetate (76.1-84.83%), linalool (9.36-10.88%) and limonene (0.87-1.80%).


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