A comprehensive technique for artificial hybridization in Chickpea (Cicer arietinum)

Abstract

BackgroundTwo crossing techniques for hybridization of chickpea have been reported and include pollination after emasculation and pollination without emasculation. Success of crossing with emasculation varied from 5 to 17%; while the success rate varied from 20 to 50% by pollination without emasculation. The important reason for the low success rate of the two procedures could be lack of detailed information on the flowering stages chosen for crossing together with the environment where plants grow.ResultsWe describe a comprehensive method for chickpea crossing where two genotypes, ICCV96029 as female and PI503023 as male parent were used. Leaf shape and seed size were used as morphological markers to select hybrids. For crossing, incision was made along the central line of the keel petal for the removal of anthers and to expose the stigma for placement of pollen from donor parent on its surface. After pollination, style was inserted back gently inside the keel petal and covered by wing petals and standard petals to make a natural sac which prevents drying of internal organs. Alternatively, if the conditions are favorable there is no need to protect the pollinated flower and therefore petal removal method for cross-pollination can be used. Our method showed around 78% crossing success rate which is much higher than the previous results.ConclusionsWe have shown that the crossing by keel petal incision or petal removal is an effective approach which significantly increases the crossing success rate. Furthermore, our detailed method shows that the flowering stage, selection of parents and temperature play crucial roles in crossing success.Electronic supplementary materialThe online version of this article (doi:10.1186/s13007-017-0202-6) contains supplementary material, which is available to authorized users.

Development of marker-free transgenic pigeon pea (Cajanus cajan) expressing a pod borer insecticidal protein

“…In our present investigation, we found that stage 2 was best for emasculation (became female) and stage 3 was appropriate for collecting pollen for crossing. Furthermore, we have also observed that stage 3 is the scientifically accurate stage for crossing 24 . The reason behind this selection is that the stigma in stage 3 showed the highest pollen acceptability, as at this stage, stigma became the most sexually active for pollination, resulting in hybridization 37 .…”

“…According to Kalve and Tadege 24 , the low success rate in the production of hybrid plants (chickpea) is due to the lack of detailed information on the flowering stages chosen for crossing. Before starting the crossing events between transgenic plants carrying cry1Ab and cre genes, we performed microscopic analysis of flowers in different stages to understand the exact timing/stage of pollination.…”

“…Stage 2 is the right time for emasculation because the anther starts to burst, so there is a chance of gating the stigma without pollination. This type of microscopic analysis of the developing reproductive organ combined with precise crossing was performed for the first time in pigeon pea, which helped to increase the crossing efficiency 24 .…”

“…This could be possible in a hybrid plant resulting from a cross between two lines. Establishment of an efficient crossing procedure is essential to create successful hybrid plants though crossing; however, it is tedious and requires specialised skills along with the knowledge of various stages of flower development 24 , 25 , 26 .…”

Genetic studies of yield components and pungency in chilli (Capsicum annuum L.) genotypes through Hayman’s approach

“…Syukur et al (2018) also stated that plant breeding is a combination of art and technology to create new superior varieties. One conventional plant breeding technique that is easy to do is artificial hybridization (Kalve and Tadege, 2017). Ganefianti et al (2019) stated that hybridization aims to combine the genetics of the two parents to produce new varieties by utilizing the heterosis phenomenon.…”