The invention relates to the field of
plant conservation and breeding optimization, in particular to a
hybrid embryo rescue method and a method for creating a large number of interspecific
hybrid seedlings of Populus nigra by using the
hybrid embryo rescue method. The method comprises the following steps: establishing an
embryo rescue
system, namely observing a separation layer between
infructescence and branches after
pollination for about 2 months,
cutting part of
infructescence to develop the
embryo rescue system, washing capsules with running water for 120 minutes, lightly wiping peels with 75%
alcohol for about 2 minutes, washing the capsules with
sterile water for 5-6 times in a super clean bench, lightly absorbing
moisture on the surfaces of the capsules with sterile
filter paper, and taking out the capsules; the method comprises the following steps of:
cutting a
capsule by using a sterile scalpel, thoroughly removing floc, carefully clamping seeds by using sterile
tweezers, putting the seeds on a 1 / 2MS culture medium containing
auxin analogues for
embryo rescue, and screening out an optimal culture medium which is 1 / 2MS + 0.06 mg / L
naphthylacetic acid (NAA) + 0.08 mg / L
indolebutyric acid (IBA) and has the pH value of 5.8-6.0 by adopting a culture dish
germination method and taking 1 / 2 MS as the culture medium.