Developing Genetic Variability of Quinoa (Chenopodium quinoa Willd.) with Gamma Radiation for Use in Breeding Programs

Luz Rayda Gomez-Pando, Ana Eguiluz-de la Barra

Cereal and Native Grain Research Program, National Agricultural University La Molina, Lima, Peru.

DOI: 10.4236/ajps.2013.42046                

Abstract

Quinoa (Chenopodium quinoa Willd.) is a staple food produced mainly by small-scale subsistence farmers in Peru’s highland. Dry seeds (cv. Pasankalla) were irradiated with doses of 150 Gy, 250 Gy and 350 Gy. In the M₁ generation, the germination process was delayed with increasing radiation dose; seedling height, root length and leaf development were most reduced at 250 Gy and at 350 Gy, no plants survived. In M₂, the maximum spectrum of chlorophyll mutations corresponded to 150 Gy and the maximum frequency to 250 Gy. The chlorine mutation was predominant, followed by xantha. Changes were registered for branch number, pedicel length, plant height, life-cycle duration, stem and foliage colour, and leaf morphology at the two doses, with improvements in plant type. More than one mutation per plant was found, especially at 250 Gy. In M₃, the same spectrum of mutations was observed, along with a valuable change in grain colour.

Keywords

Chenopodium quinoa; Gamma Rays; Mutant

1. Introduction

Quinoa (Chenopodium quinoa Willd.) is native crop of the Andes. Quinoa is cultivated in Peru by small farmers in plots below of 1 ha [1]. Its tolerance to drought, frost and saline soils makes it important in highland farming systems [2,3]. This annual crop belongs to the Chenopodiaceae (goosefoot) family. Quinoa has an exceptionally nutritious balance of protein, fat, oil and starch. Grains average 16% protein, higher than common cereals. Moreover, the protein is of unusually high quality, and is close to the FAO standard for human nutrition. Quinoa protein is high in the essential amino acids lysine, methionine and cystine, making it complementary to both other grains (deficient in lysine) and legumes (deficient in methionine and cystine) [4-6].

Quinoa is planted mostly in Puno to altitudes of over 3800 m. Due to the stressful climatic conditions and the technology in use there, productivity is low; for many years it ranged from 900 to 1100 kg/ha [1]. There are many ways to improve crop performance: one of them is genetic improvement via mutation induction. According to the IAEA database, there are more than 2500 mutant varieties of more than 170 different species [7]. There are many reports of improved morphological and physiological characteristics in cereals, grain legumes, oil seedsfiber crops, vegetables and ornamentals following mutation induction, with gamma rays being the preferred agent, and plant type and yield are the traits most commonly reported [8-10]. Although the quality of many crops has been improved thru mutation induction [11], to the best of our knowledge, this technique has not been applied in quinoa to improve its agricultural performance in the marginal conditions of the Peruvian highlands but it was applied in barley and Amaranthus [12].

The Cereal and Native Grain Research Program at the National Agricultural University La Molina (Peru) is making efforts to improve the early cultivar PasankallaINIA, which has a growing period of 140 days; the plant can measure up to 120 cm in height; its panicle is light purple, and it has a glomerular inflorescence of intermediate density. Seeds have a reddish-brown pericarp and the episperm (seed coat) is lead-colored.

2. Materials and Methods

2.1. Plant Material

Two hundred grams of dry quinoa seeds with 12% of moisture of cv. Pasankalla were irradiated with gamma rays at the Peruvian Institute of Nuclear Energy (IPEN), at doses of 150, 250 and 350 Gy. Non-irradiated seeds were used as controls in each evaluation. Seeds have to be delivered for irradiation to a nuclear centre. Such centres have been established in most countries or at IAEA Headquarters in Vienna.

2.2. Percent of Germination in M₁ Generation

To establish percent of germination, 100 seeds per Petri dish (Ø: 10 cm, height: 2 cm) were placed on filter paper and covered with tissue paper. Each treatment included four Petri dishes (replicates) (for a total of 400 seeds per treatment). Distilled water (5 ml) was added at the beginning of the experiment. Every dish was then irrigated daily with 1 ml water to maintain moisture. The experiment was conducted under a photosynthetic photon flux density of 458 µmol/m²s (12 h/day) at 20˚C. The germination percentage was recorded on days 3, 5, 7, 9, 11 and 15.

2.3. Physiological Damage in M₁ Generation

To determine the physiological damage or somatic effects caused by the irradiation, seedling height and root length were measured in 1-month-old plants using the “sandwich blotter” technique [17]. Thirty seeds from each treatment were used in this experiment in four replicates (for a total 120 seeds per treatment). The level of somatic effects after mutagenic treatment can be evaluated on the basis of various parameters, including delay in seed germination; level of disturbances in the cell cycle; reduced seedling emergence; reduced seedling and plant growth; appearance of chlorophyll defects; and reduced fertility and plant survival. The term “reduced” indicates change in expression of a particular character in relation to the control, usually the parent cultivar or the breeding line whose seeds were treated with a mutagen [14]. The rest of the seeds from each treatment were sown in beds under field conditions following the protocol described for quinoa crop in Peru [15]. The formation of cotyledonal and true leaves and percent of survival were evaluated. Individual heads or inflorescences of the M₁ generation were harvested. Data were analyzed using a completely randomized design.

2.4. Evaluation of M₂ Generation

The M₂ population was cultivated in the field, one inflorescence per row. The following traits were evaluated: chlorophyll mutations, life-cycle duration, plant height, and morphological characteristics. Putative mutants were identified and harvested separately as individual plants. This generation was cultivated following the protocol described for different crops under isolated field conditions [14].

2.5. Evaluation of M₃ Generation

The selected putative M₂ mutants were cultivated as row/ plant (in the field, one plant per row) for progeny determination and homozygosity testing. This generation was cultivated following the protocol described for different crops under field conditions planted in a row or rows depending on the amount of seed [14].

2.6. General Experimental Scheme

Figure 1 shows the experimental scheme followed in this research.

Conflicts of Interest

The authors declare no conflicts of interest.

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